Collusion

Collusion is an agreement between two or more parties, sometimes illegal and therefore secretive, to limit open competition by deceiving, misleading, or defrauding others of their legal rights, or to obtain an objective forbidden by law typically by defrauding or gaining an unfair advantage. [citation needed] It is an agreement among firms or individuals to divide a market, set prices, limit production or limit opportunities. [1] It can involve â€Å"wage fixing, kickbacks, or misrepresenting the independence of the relationship between the colluding parties†. 2] In legal terms, all acts affected by collusion are considered void. [3] | In the study of economics and market competition, collusion takes place within an industry when rival companies cooperate for their mutual benefit. Collusion most often takes place within the market structure of oligopoly, where the decision of a few firms to collude can significantly impact the market as a whole. Cartels are a special case of e xplicit collusion. Collusion which is not overt, on the other hand, is known as tacit collusion. How is OPEC a collusive oligopoly? Answer:OPEC is a collection of oil exporting countries. Oligopoly – Industry that is controlled by a few major players (firms or countries) Collusion – When industry leaders secretly agree to limit quantities of production. This will guarantee the colluders a higher price for their product OPEC meet to discuss the quantity of oil they will allow onto the world market. This is collusion. Because the OPEC members are the main suppliers of oil they are said to be an Collusion and Cartels by David A. Mayer One of the blessings of competition is that it leads to lower prices for consumers.For the producer, however, this blessing is a curse. Low prices often mean low profits. Given a choice between competition and cooperation, profit-maximizing firms would more often than not prefer cooperation. Regardless of what you learned in kindergarten, yo u do not want the businesses you buy from to cooperate. You want them to compete. Adam Smith, the father of modern capitalism, warned that nothing beneficial comes from the heads of business getting together. In the United States, firms are forbidden from cooperating to set prices or production.The abuses of the late nineteenth and early twentieth century trusts were the impetus for the â€Å"trust-busting† of President Theodore Roosevelt. With the Sherman Antitrust Act and later the Clayton Antitrust Act, the government prohibited outright collusion and other business practices that reduced competition. Prior to OPEC, world oil prices were mainly under the control of the Texas Railroad Commission. With the rise of OPEC came a shift in power from U. S. producers to the oil states of the Middle East. Even though it violates the law, businesses from time to time will collude in order to set prices.Colluding firms can divide up the market in a way that is beneficial for them. Th e firms avoid competition, set higher prices, and reduce their operating costs. Because collusion is illegal and punishable by fine and prison, executives at firms are reluctant to engage in the practice. The meetings of business leaders are almost always in the presence of attorneys in order to avoid the accusation of collusion. Forming Cartels Businesses that collude may form cartels. A cartel is a group of businesses that effectively function as a single producer or monopoly able to charge whatever price the market will bear.Probably the best-known modern cartel is the Organization of the Petroleum Exporting Countries, or OPEC. OPEC is made up of thirteen oil-exporting countries and is thus not subject to the antitrust laws of the United States. OPEC seeks to maintain high oil prices and profits for their members by restricting output. Each member of the cartel agrees to a production quota that will eventually reduce overall output and increase prices. OPEC is bad news for anyone that enjoys cheap gasoline. Fortunately for consumers, cartels have an Achilles heel.The individual members of a cartel have an incentive to cheat on their agreement. Cartels go through periods of cooperation and competition. When prices and profits are low, the members of the cartel have an incentive to cooperate and limit production. It is the cartel's success that brings the incentive to cheat. If the cartel is successful, the market price of the commodity will rise. Individual members driven by their own self-interest will have an incentive, the law of supply, to ever-so-slightly exceed their production quota and sell the excess at the now higher price.The problem is that all members have this incentive and the result is that eventually prices will fall as they collectively cheat on the production quota. Cartels must find ways to discourage cheating. Drug cartels use assassination and kidnapping, but OPEC uses something a little more civilized. The single largest producer in th e cartel is Saudi Arabia. Saudi Arabia also has the lowest cost of production. If a member or members cheat on the cartel, then Saudi Arabia can discipline the group by unleashing its vast oil reserves, undercutting other countries' prices, and still remain profitable.After a few months or even years of losses, the other countries would then have an incentive to cooperate and limit production once again. * Definition: OPEC stands for The Organization of Petroleum Exporting Countries. It is an organization of 12 oil-producing countries that effectively control the world's oil. OPEC members pump out 42% of the world's annual supply, controlling 61% of exports. This situation isn't likely to change, since these 12 countries hold 80% of the world's proven oil reserves. For these reasons, OPEC's decisions are critical to countries that depend on oil imports.What Does OPEC Do? OPEC states quite plainly that its goal is to manage the world's supply of oil. It does this to make sure its mem bers get what they consider a good price for their oil. Since oil is a fairly uniform commodity, most of its consumers base their buying decisions on nothing other than price. What's a good price? In the past, OPEC said it was around $70-$80 per barrel. If prices drop below that target, OPEC members agree to restrict supply to send prices higher. Otherwise, they would wind up increasing the supply to make more national revenue.By competing with each other, they would drive prices even lower. This would stimulate even more demand, and OPEC countries will run out of their most precious resource that much faster. When prices are higher than $80 a barrel, oil-producing countries would naturally want to produce more to bring in extra national revenue. However, if they did that, they increase supply, lowering the price. Instead, OPEC members agree to produce only enough to keep the price high for all members. Furthermore, if prices are too much higher than $80 a barrel, then other countri es have the incentive to drill more expensive oil fields.Sure enough, now that oil prices are closer to $100 a barrel, it's become cost effective for Canada to explore its shale oil fields, and for the U. S. to use fracking. As a result, non-OPEC supply has increased. OPEC's second goal is to reduce oil price volatility. That's because, at current prices and rates of production, OPEC countries have enough oil to last for 113 years. In addition, oil is expensive to produce. For maximum efficiency, oil extraction must run 24 hours a day, seven days a week. In addition, closing facilities could physically damage oil installations and even the fields themselves.Ocean drilling is especially difficult and expensive to shut down. Therefore, it's in OPEC's best interests to keep world prices stable. For example, in June 2008, prices spiked to $143/barrel. OPEC responded by agreeing to produce a little more oil, which brought prices down. However, the global financial crisis brought oil pric es down to $33. 73/barrel in December. OPEC responded by reducing the supply, helping prices to again stabilize. A slight modification is usually enough to restore price stability. OPEC also adjusts the world's oil supply in response to crises and shortages.For example, it replaced the oil lost during the Gulf Crisis in 1990. Several million barrels of oil per day were cut off when Saddam Hussein armies destroyed refineries in Kuwait. OPEC alos increased production in 2011 during the crisis in Libya. The Oil and Energy Ministers from the OPEC members meet twice a year, or more if needed, to coordinate their oil production policies. Each member country abides by an honor system, agreeing to only produce a certain amount. However, if a country winds up producing more, there really is no sanction or penalty.Furthermore, each country is responsible for reporting its own production. Therefore, there is room for â€Å"cheating. † On the other hand, a country won't go too far over i ts quota, since it doesn't want to risk being kicked out of OPEC. Despite its power, OPEC cannot completely control the price of oil. In some countries, additional taxes are imposed on gasoline and other oil-based end products to promote conservation. More importantly, oil prices are actually set by the oil futures market. Much of the oil price is determined by these commodities traders. For more on this, see Why Are Oil Prices So High?OPEC Members OPEC members are Algeria, Angola, Ecuador, Iran, Iraq, Kuwait, Libya, Nigeria, Qatar, Saudi Arabia, UAE and Venezuela. Saudi Arabia alone produces enough oil to materially impact the world's supply. For this reason, it really has more authority and influence than the other countries. Here's a ranking of production by member: 1. Saudi Arabia – 9. 311 million barrels/day. 2. Iran – 3. 576 mb/d. 3. Venezuela – 2. 881 mb/d. 4. Kuwait – 2. 659 mb/d. 5. Iraq – 2. 653 mb/d. 6. UAE – 2. 565 mb/d. 7. Niger ia – 1. 975 mb/d. 8. Angola – 1. 618 mb/d. 9. Algeria – 1. 162 mb/d. 10. Qatar – . 734 mb/d. 11.Ecuador – . 5 mb/d. 12. Libya – . 489 mb/d. (Source: OPEC Annual Statistical Bulletin 2012) Many non-OPEC members also voluntarily adjust their oil production in response to OPEC's decisions. In the 1990s, they learned that increasing their own production to take advantage of OPEC's restraints meant oil prices stayed low, restricting profits for everyone. These cooperating non-OPEC members include Mexico, Norway, Oman and Russia. OPEC History In 1960, five OPEC countries formed an alliance to regulate the supply, and to some extent, the price of oil. These countries realized they had a non-renewable resource.If they competed with each other, the price of oil would be so low that they would run out sooner than if oil prices were higher. This first meeting was held September 10-14 1960 in Baghdad, Iraq. The five founding members were Iran, Iraq, Kuw ait, Saudi Arabia and Venezuela. OPEC was registered with the UN on November 6, 1962. (Source: OPEC Frequently Asked Questions) Article updated March 13, 2013 The acronym OPEC is short for the Organization of Petroleum Exporting Countries. Through its 12 member countries, the group controls nearly 80% of the world’s crude oil reserves and about 45% of its worldwide production.This makes it extremely influential in the market for crude oil and its derivatives, like gasoline and diesel fuels. OPEC member countries include: Algeria, Angola, Ecuador, Iran, Iraq, Kuwait, Libya, Nigeria, Qatar, Saudi Arabia, United Arab Emirates and Venezuela. Most of these countries rely on oil prices to sustain their 408 million combined inhabitants. Consequently, OPEC was designed to unify petroleum policies, ensure price stability and facilitate market efficiency. The Origins of OPEC OPEC was originally started by Iran, Iraq, Kuwait, Saudi Arabia and Venezuela in the 1960s.While their initial o bjectives were reasonable, OPEC was soon labeled a cartel by many Western countries, given its practice of adopting output rationing in order to maintain certain price levels. Similarly, its political motivations have also contributed to its image. In 1973, OPEC became infamous for its oil embargo that punished the United States and Western Europe for its support of Israel against Arab nations in the Yom Kippur War. The resulting shortage led to limits on gas available at gas stations and eventually a worldwide economic recession that saw high unemployment and inflation.OPEC’s Declining Influence In the 1980s, OPEC suffered from a six-year decline in oil prices due to reduced demand and over-supply that led to a glut in the world market. While OPEC lost its unity during the 1980s and early 1990s during the Gulf War, oil prices recovered after the September 11th attacks against the United States and the subsequent invasion of Afghanistan. As of 2011, OPEC continues to publish recommended production quotas designed to increase the price of oil, but member countries aren’t as keen as they used to be on compliance.For instance, one Saudi Arabian official told the New York Times that the country would meet the market’s demand, presumably despite any quotas from OPEC. OPEC Finally Reaches an Agreement In December of 2011, OPEC reached a new agreement on production quotas for the first time in three years. Output from the 12 member states was set at 30 million barrels per day, which was roughly in-line with the supply at the time. The result in the financial markets after the announcement was a muted 1. 8% decline in crude oil futures.OPEC leaders also discussed how to handle the decreased production from Libya after the fall of Moammar Qaddhafi. Shortly after the supply cut, Saudi Arabia stepped in and increased production, which was met with distaste from Iran and Venezuela. The matter was resolved by temporarily eliminating country-specific qu otas until a June 2012 meeting. OPEC’s Influence on Investors While OPEC hadn’t agreed on production targets for some time, primarily due to Saudi Arabia’s opposition, the organization did manage to set a new production target in late-2011 of 30 million barrels per day, which was largely in-line with current production.The agreement could mean more cohesion among its membership and additional quotas imposed down the line. Those trading in the crude oil or derivative markets since the 1970s are very familiar with OPEC’s influence on pricing. If the organization can agree on set prices, their control over a large portion of the market enables them to significantly influence prices. Until 2011, this was largely a non-issue given Saudi Arabia’s opposition to any limits. Key Takeaway Points * OPEC began in the 1960s as a way to control oil prices in countries where oil was a primary source of livelihood for citizens. OPEC turned political in the 1970s a fter the oil embargo and member disagreements hurt its power throughout the 1980s and early 1990s. * While OPEC has struggled with member cohesion, the organization has managed to strike an agreement in late-2011 and will meet again in June of 2012. Definition Collusion occurs when firms in a market chose an optimal level of output for that market in order to maximize total industry profits (Baye, 2006). Collusion typically occurs in the oligopoly market model when the number of firms are few as opposed to many.This can simply be explained by the fact that when firms are working together, the more firms that have to work together the harder it is to make everyone happy. See Wiki page about oligopoly for further information: http://mbaecon. wikispaces. com/oligopoly%26nbsp%3B. See Wiki page about monopolistic competition to see why collusion is more difficult with many firms: http://mbaecon. wikispaces. com/monopolistic+competition. Monitoring the agreement This can be seen in that f irms must monitor one another such that their collusion agreement is kept.This can be accomplished by monitoring the other firms in the collusion agreement. There comes a point at which because there are so many firms in the agreement that the costs of monitoring the other firms outweighs the benefits from the collusion agreement. This monitoring can be seen by the formula n*(n-1) where n is the number of firms in the agreement. If there are six firms in agreement there must be 30 (6*(6-1)=30) monitors to keep everyone aware that all the firms in the agreement are holding to the terms. As the number of firms increase in the market the number of monitors increases dramatically.How collusion occurs The first way collusion occurs is that firms will meet and agree not to steal each others customers, and if one firm tries to steal anothers customers there will be retaliation. This form of collusion is called explicit collusion. Another way firms collude may not involve physically meeting or talking at all. Overtime firms may reach a nonverbal understanding that they will leave each other alone, but if one firms tries to steal customers there are consequences. The second form of collusion just discussed is called tacit collusion.Tacit collusion occurs when the behaviors of the players in the market are learned. If you try to steal customers and get attacked back, eventually your firm will probably stop trying to steal customers. On the other hand if you lower prices in order to steal customers and there is no retaliation, or the retaliation is not effective, tacit collusion will not occur. Legality Collusion is considered illegal within the United States, European Union, and Canada. Collusion falls within the category of antitrust laws/competition laws. These are laws that prohibit anti-competitive behavior and unfair business practices.These laws make certain practices illegal because they hurt the businesses, consumers, or both, typically violating standards of et hical behavior (wikipedia-antitrust, 2006). Tacit collusion because of the fact that it is the learned behaviors of the players in the market is much more difficult to enforce, because specifically there has been no formal agreement, because of this tacit collusion can and does occur today. Questions: Which is not a form of collusion? A. ) The behaviors of a competing firm in the same market of a second firm are learned. B. Two firms meet and agree not to steal one another's customers. C. ) If an agreement has been reached to not steal one another's customer has been breached, retaliation will occur. D. ) One firm lowers prices to compete against another when there was no agreement against it. Answer: D. ) This is just the normal game of business that occurs every day. There is no collusion because one firm is lowering a price without specific knowledge or an understanding that there will be repercussions or action taken or not taken because of this action. In a finite number of gam es collusion will be more likely to occur: A. On the second to last turn. B. ) From the beginning. C. ) Once a tacit understanding of business practices has been reached. D. ) Will not occur because there is no effective punishment method that can be used. Answer: D. ) Will not occur because there is no effective punishment method that can be used. This is because of the ending nature of the finite number of games to be played. Each period the players in the game know what the last period will hold, and because they know the last periods outcome, the second to last game is the last game.It is because the second to last game is now the last game that once again the players know how each member of the collusion will act because there is no punishment that can be effective. This continues on until the first game being played, and each member of the collusion knows that each member will cheat, so collusion will not occur. Sustained collusion is more likely to occur when firms know: A. ) their rivals. B. ) who their rivals customers are. C. ) when their rivals deviate from the agreement. D. ) All the above. Answer: D. ) All the above.This are all reasons why a sustained collusion is more likely to occur. The last reason not listed is that firms must be able to successfully punish rivals for deviating from the agreement. A small firm with 1 outlet and a large firm with 10 outlets decide to collude, the small firm: A. ) is at an advantage because they only have to focus on the big competitor. B. ) is at an advantage because they are now â€Å"safe† from the big competitor. C. ) is at a disadvantage because they have to monitor more locations then the larger firm does. D. is at a disadvantage because they have less bargaining power when the â€Å"contract† needs to be renegotiated. Answer: C. ) is at a disadvantage because they have to monitor more locations then the larger firm does. This is because economies of scale exist within the monitoring act. Th e larger firm only has to monitor the one outlet of its collusive partner. The smaller firm has to monitor the larger firms 10 outlets, which most likely will cost more and be a larger percentage of the â€Å"savings† associated with the collusive agreement.

Hyperion Financial Management: Performance Tuning Guide

For comprehensive tuning information for the PM System, please refer to the [email  protected] Hyperfine PM, Fusion Edition Performance and Tuning Guide. Top of Document This document is written for people who monitor performance and tune the components in an PM/BI environment. It is assumed that readers know server administration and hardware performance tuning fundamentals, web servers, java application servers and database. Introduction to Oracle Hyperfine PM System Performance To maximize Oracle Hyperfine PM System performance, you need to monitor, analyze, and tune all the components.This guide describes the tools that you can use to monitor performance and the techniques for optimizing the performance of Oracle Hyperfine PM System components, for example Financial Management, Sybase, Reporting and Analysis and Planning. Performance Terminology This guide uses the following performance terms: Scalability The system's ability to perform within specification under increasing use r load, data load and hardware expansion. Latency The time between the issuing of a request and the time when the work actually begins on the request. Think time The time a real user pauses to think between actions.Resource utilization A consumption metric, for example, the percent of CPU usage. Response time A time metric, for example round-trip time it takes the server to deliver a Web page. Throughput A rate metric (requests per unit of time), for example, requests per second, bits per second. For example, if an application can handle 20 customer quests simultaneously and each request takes one second to process, this site has a potential throughput of 20 requests per second. Understanding Key Performance Drivers To optimize your deployment, you must understand the elements that influence performance and scalability.A factor that dictates performance is called a key performance driver. Knowing how the drivers behave in combination further enhances your ability to deploy Oracle Hy perfine PM system optimally, based on the unique requirements of each deployment. Hardware Capacity – Factors such as number of servers, quantity and speed of processors, available RAM, network speed etc. Technical Platforms Tuning – Fine tuning other third party software required for installing and running Oracle Hyperfine PM products; for example: relational databases, Java application servers, Web servers, Server / Client Operating System and browsers.Business Application Design – Application design is an important factor in system performance I. E. Structure, size, and use of product features in designing applications' databases, reports, Web data entry forms, calculations and consolidations. Business Process usage – Activities carried out by users in the normal flow of your business cycle. Business process usage has three components: User activity-?Activities available to users for data load or data entry, database processing (consolidations, copy, c lear, and so on), and reporting and analysis. Rate of user activity – A number of transactions executed by one user per one hour.User concurrency-?Number of users for each activity being carried out simultaneously. Tuning Recommendations for Financial Management Performance tuning Oracle Hyperfine Financial Management is a complex and iterative process. To ghetto started, this document includes recommendations to help you optimize your Oracle Hyperfine HEM system reference. Note: tuning has to be done for a particular production workload. Tuning can be conducted when workload is generated by load generation tools like Oracle Application Testing Suite (TATS) or Load Runner by HP.This document touches on several areas that provide a quick start for performance tuning Financial Management, including: Tune Operating Systems parameters Tune HTTP Server parameters Tune HTTP Server Compression / Caching 4 Tune Oracle Database Parameters Hyper-Threading / SMS Considerations Note: whi le the list in each of the above stated section is a useful tool in tarring your performance tuning, it is not meant to be comprehensive list of areas to tune. You must monitor and track specific performance issues within your implementation to understand where tuning can improve performance.Top Of Document Diagnosing Performance Problems When a performance issue arises, it is critical to first determine the cause prior to taking any corrective action. Oracle does not recommend changing performance-related parameter settings or taking other actions until an extensive analysis of the problem has been performed. Using Monitoring Tools Oracle strongly recommends using monitoring tools to collect performance ATA as part of the diagnostic process. Monitoring the WFM application server, web servers, database server(s) and network layers provide useful performance data.The recommended tool to monitor the WFM application process performance on the Windows server side is Microsoft Performanc e monitoring. Steps for configuring Performance Monitor to gather the counters specific for HEM application can be found here: HTTPS://support. Us. Oracle. Com/pop/faces/secure/km/ Documentation's. JSP? Id=953294. 1 Performance counters to monitor include: Us beseem Counter Guidance Memory Memory: Free System â€Å"Warning† when Free System Page Table Entries is Page Table Entries less than 8,000 â€Å"Critical† when Free System page Table Entries is less than 5,000.On WFM systems if you enable KGB without proper tuning as per recommendations stated in section â€Å"Tune Windows /KGB†, you will notice 5000 PET as initial value after the reboot. Memory: Available Should be no lower than 20% to 25% of Installed Maybes physical memory. In these cases, carefully monitor Paging activity. Memory: Page This counter should be below 1,000 at all times. Reads/sec Processor processor: % processor Total processor utilization should be lower than about Time Total to 80%. Pro cessor: % Processor Each processor instance should be lower than about 5 Disk 70% to 80% utilization.

Functional Immunity

This type of immunity arises from  customary international law  and  treaty law  and confers immunities on those performing acts of state (usually a foreign official). Any person who in performing an act of state commits a criminal offence is immune from prosecution. This is so even after the person ceases to perform acts of state. Thus it is a type of immunity limited in the acts to which it attaches (acts of state) but will only end if the state itself ceases to exist.This type of immunity is based on respect for sovereign equality and state dignity. The offices usually recognised as attracting this immunity are Head of State or Head of Government, senior cabinet members, Foreign Minister, and Minister for Defence: see the  Arrest Warrant Case,Pinochet Case  (R v Bow Street Magistrates; ex parte Pinochet Ugarte (No 3)  [2000] 1 AC 147, House of Lords).Such officers are immune from prosecution for everything they do during their time in office. For example, an English court held that a warrant could not be issued for the arrest of  Robert Mugabe  on charges of international crimes on the basis that he was a presently serving Head of State at the time the proceedings were brought:  Mugabe, reported at (2004) 53 ICLQ 789. Other examples are the attempts to prosecute  Fidel Castro  in Spain and  Jiang Zemin  in the USA.However, the moment accused leaves office, they are liable to be prosecuted for crimes committed before or after their term in office, or for crimes committed whilst in office in a personal capacity (subject to jurisdictional requirements and local law). Pinochet  was only able to come to trial because Chile and the UK had both signed and ratified the UN Convention Against Torture through which such immunities were waived. It may be the case that personal immunity is itself being eroded.In 2004 the Appeals Chamber of the  Special Court for Sierra Leone  held that indicted Liberian president  Charles Taylor  cou ld not invoke his Head of State immunity to resist the charges against him, even though he was an incumbent Head of State at the time of his indictment. However, this reasoning was based on the construction of the court's constituent statute, that dealt with the matter of indicting state officials. In any case, Taylor had ceased to be an incumbent Head of State by the time of the court's decision so the arresting authorities would have een free to issue a fresh warrant had the initial warrant been overturned. Nevertheless, this decision may signal a changing direction in international law on this issue. Recent developments in international law suggest that this type of immunity, whilst it may be available as a defence to prosecution for local or domestic crimes or civil liability, is not a defence to an international crime. (International crimes include  crimes against humanity,  war crimes, and  genocide).This has developed in the jurisprudence of the International Criminal T ribunal for the Former Yugoslavia, particularly in the  Karadzic,  Milosevic, and  Furundzija  cases (though care should be taken when considering ICTY jurisprudence due to its  Ad-hoc  nature). This was also the agreed position as between the parties in their pleadings in the  International Court of Justice  Case Concerning the Arrest Warrant of 11 April 2000 (Democratic Republic of the Congo v. Belgium).The reasons commonly given for why this immunity is not available as a defense to international crimes is straight forward: (1) that is genocide, war crimes and crimes against humanity are not acts of state. Criminal acts of the type in question are committed by human actors, not states; and (2) we cannot allow the  jus cogens  nature of international crimes, i. e. the fact that they are  non-derogable  norms, to be eroded by immunities. However, the final judgment of the ICJ regarding immunity may have thrown the existence of such a rule limiting functional immunities into doubt.See in this respect the criticism of the ICJ's approach by Wouters, Cassese and Wirth among others, though some such as Bassiouni claim that the ICJ affirmed the existence of the rule. Regarding claims based on the idea that a senior state official committing International crimes can never be said to be acting officially, as Wouters notes â€Å"This argument, however, is not waterproof since it ignores the sad reality that in most cases those crimes are precisely committed by or with the support of high-ranking officials as part of a state’s policy, and thus can fall within the scope of official acts. Academic opinion on the matter is divided and indeed only the future development of International Customary law, possibly accelerated by states exercising  universal jurisdiction  over retired senior state officials, will be able to confirm whether state sovereignty has now yielded partially to internationally held human rights values.In November 2007 , French prosecutors refused to press charges against former US Secretary of Defense Donald Rumsfeld for torture and other alleged crimes committed during the course of the US invasion of Iraq, on the grounds that heads of state enjoyed official immunity under customary international law, and they further claimed that the immunity exists after the official has left office. [1]

Compares the significance of the dominant conservative parties and Essay

Compares the significance of the dominant conservative parties and their opponents on the left (and in the center, as in the case of the DPJ) in France and Japan - Essay Example Due to this unity, the Japan Democratic Party won the next elections, held this power for over almost five decades, but lost the power in 1993. The party strategies came from its strong ties with the United Nation and the USSR. However, Criminal Intelligence Agency in US was not happy with the Socialist and the Communist party and therefore spent millions of dollars influencing Japans elections in the favor of the Liberal Democratic Party. Many analysts argue that the influence was due to the second world war that really affected the Japanese hence the CIA were worried of their counter reaction. The influence of the US CIA was a deep kept secret and was only revealed later in the 1990 thanks to The New York Times. In the 1960’s LDP managed to host 1964 Olympics and neutralize the Vietnam War (Fackler, Osaka Mayors Radical Message Has Broad Appeal in a Weary Japan). Their success continued as the Party managed to improve the country’s economy. However, the Party did not have it smooth all the way. The Party encountered resistance from the Socialist Party and Japan Community party. LDP also encountered many scandals including some of young members breaking away from the government and joining their opponents. Despite this, the government managed to consolidate and move forward. The Party made policies from all specters of the economy that managed to hold the party into leadership until 1993 when it lost. The party lost due to lack of power to influence voters in addition to many scandals in the government. Despite the loss the LDP was able to gunner its power back and after three years it was back to leadership. In 1998, Democratic Party of Japan was formed. The party challenged LDP mostly in 2003 and 2004 in parliamentary elections (Sieg). France political interest began hundreds of year ago with philosophers such as Locke challenging the government in its governance role.

Marketing Management Essay Example | Topics and Well Written Essays - 2500 words - 1

Marketing Management - Essay Example Moreover, the promotions presented by the management of Coca-Cola for the drinks are also quite fascinating and effective for the individual that acted as a boon for the growth and development of the organization in the market of North Korea as compared to other rival players (The Coca-Cola Company, 2013). Customer Analysis From the below presented graph, it might be clearly described that, Coca-Cola mainly targets the young generation of the country of North Korea with extreme consciousness for the health and hygiene. Moreover, the refreshing drink of Coca-Cola is mainly preferred by the age groups within 15-35 years among others as it serves as the best companion in parties, festivals, occasions and many others. And also the drink might be consumed by the individual of high to medium income groups along with a medium life style. Thus, it might be depicted that the drinks of Coca-Cola is mainly targeted to all the individual of the society with a proposition of ‘Healthy and re freshing drink’ among other rival players (The Coca-Cola Company, 2013). Objective and Strategies The prime objective of the organization of Coca-Cola is to offer best quality of drinks to the customers so as to retain its prosperity and loyalty within their minds. Apart from this, the organization of Coca-Cola also desires to offer most advantageous and refreshing drink so as to make it a value-added product in the market among other rival products. Only then, the desire to enhance the brand image and reputation of the organization might get enhanced and effective in future era. And the market share and profit margin of the organization might also get enhanced to almost 25% as compared to other rivals (The Coca-Cola Company, 2013). In order to make the objectives of the organization successful, the management might desire to implement varied types of strategies such as competitive growth policies and expansion survival strategies. These strategies might prove effective for t he organization thereby amplifying its brand value and market growth to a significant extent as compared to others. Along with this, by implementing expansion survival strategy, the organization might expand its products thereby improving the statistics of the organization in the market of North Korea. And it might also prove helpful in amplifying the brand value and market share in North Korean market (The Coca-Cola Company, 2013). Corporate Objectives The most essential requirement of any organization operating in this age is sustainability. This is because; it might help an organization to tackle varied types of challenging situations in an effective way as compared to many other existing rival players. Some of the objectives are stated below: Profit- presenting maximum extent of the returns to the shareholders thereby enhancing its foothold in the market of North Korea. Human resource- trained and experienced human resources need to be recruited within the organization so as to enhance the portfolio and reputation in North Korean m

FEDEX REPORT Essay Example | Topics and Well Written Essays - 1250 words

FEDEX REPORT - Essay Example As far as product and market destinations are concerned, FedEx is known to offer the second largest civil fleet service across the United States and other Caribbean Islands including Puerto Rico and Dominican Republic (Breiman et al, 2009). There also is FedEx Ground, which is a dedicated courier service for Canada and other North American countries. FedEx also offers customer specification and tailor services including home delivery, custom and insurance provided through FedEx Trade Network, supply chain service, corporate services, print service, as well as FedEx Ship Centers. As a way of ensuring that its products meet customer specification and needs, there is a dedicated FedEx product named FedEx TechConnect, which offers toll free customer support to customers all around the globe. It has been said that through FedEx TechConect, FedEx creates a competitive advantage of learning from within its own rank, why and how it will succeed with its product strategy. Pricing Strategies P ricing has been said to be a very important marketing strategy for the creation of competitive advantage in a globally competing market. ... The price discount on FedEx Ground is one of such popular pricing strategies used by the company to ensure that customers get value for money. What is more, FedEx tries to keep its traditional base cost of service as minimal as possible. For example pick up services are offered by FedEx at a traditional rate of $6 per delivery (Carter, Daniel and Betty, 2006). In relation to key competitors on the American and Canadian market, this traditional cost has been said to be one of the lowest. In addition to the traditional cost offered on pick up services, there is an additional weekly charge of $6 for business locations that make requested pick up services. Placement Strategies In terms of location, FedEx is predominately situated in the United States and Canada. These are where most of its stationery offices are. But thanks to globalization and flight services, this limited placement is not in any way a limitation to the distribution strategy adapted by the company. This is because in it s scheme of distribution strategy, the company operates on a borderless strategy whereby it opens up its service to all continents of the world. To make this strategy sustainable, FedEx has an air fleet service dedicated for an inter-continental goods and mail delivery. Even though the company reports of effectiveness with this distribution strategy, experts have said that FedEx could reach a more deepened mileage with its products and services if the company expanded its placement by way of opening more offices and centers across its key regional markets (Icove, Karl and Vonstorch, 2005). It has been suggested for example that the company had continental head quarters, from which it would operate a more decentralized placement strategy. The advantage of such a decentralized placement

The four parts of the S.A.R.A. problem-solving process Essay

The four parts of the S.A.R.A. problem-solving process - Essay Example Analysis is the use of different informational sources to find out the cause of a problem, the responsible parties for it, the entities that are affected, the location of the problem, it’s time of occurrence, and the form it takes. It requires identification of the patterns which describe the conditions facilitating the problem. Response means the execution of actions that not only address the problem analysis phase’s most important findings but also address at least two of these; deflecting offenders to prevent occurrences of problem again, safeguarding the likely victims, and reducing the tendency of crime locations to be problematic. Assessment means gauging the responses’ impact on the targeted crime problem by use of the collected information from different sources before as well as after their implementation. An example of its use by an officer is when a problem like a murder needs to be investigated in a neighborhood. The S.A.R.A. model helps the officer identify the problem’s cause, its stakeholders, effectively respond to it, and assessing its impacts to ensure it does not happen

J.S. Bach's Cantata 91 Essay Example | Topics and Well Written Essays - 1000 words

J.S. Bach's Cantata 91 - Essay Example It will be shown how Bach used the different orchestration techniques to illustrate the differences of emotion and the development of the life of Jesus. There are the use of voices, instrumentation and the use of the words of the Cantata to bring this beautiful piece together. The words of the Cantata were taken from the writings of Martin Luther. In the 16th century, he based his liturgical melodies partly on melodies from the middle ages in the 14th century. It is not known who wrote some of the hymn. One must note that Germany was not a Catholic country. The leise or (hymn) "Gelobet seist du Jesu Christ" was used to replace psalm chorals before sermons at Christmas time.(www.bach-cantatas.com So it was an appropriate title for Bach to use. Bach uses three of the verses. "Jesus was born as a human being and became the Son of God. He is eternal as he is humble as he was born from flesh and blood. We should thank him eternally." He adds an additional 2 verses at the beginning and end (2,3,2). The cantata tells a story of Jesus birth thus the Christmas Cantata. There are 6 movements but 7 verses. Each movement is attached with the use of Kyrie Eleison as the last phrase. The orchestration is typical for his Cantatas - chorus, strings, 3 oboes, 1 bassoon, 3 trumpets, continuous and 1 oboe de cacia) The use of trumpets and oboes was to give contrast to the voices in color, texture and tonal variation. The beginning is a time of praise and joy with the birth of Jesus. In the Overture, there is an long instrumental introduction. The opening choral sings "Gelobet seist du Jesu Christ". The instrumentation introduction is intertwined with the choral. This differs from the heavy choruses of composers like Handel in the Messiah. The horns intertwine with the strings and the voices to show the joy of the birth and Mary and the angels. The choral sings a three note motif which is then used to unite the Cantata The first part as other verses end with the common use of Kyri e Eleison or in German Kirieleis (Our Lord). The 2 techniques are used to join the movements: the use of the Kyrie Eleison and the second point is the music technique of using a motif which is first introduced by the choral in the first verse and then used to join each of the verses. In line 4 and 5, it is used by the choral. The second movement is sung with a soprano recitative answered by a soprano choral (4 out of the 5 phrases) showing Jesus, the son of God is still in bodily form. The strings are used with the continuous only. Bach joins the last phrase with the word Kirieleis to be sung. Jesus has divine power. In the composition, accompanied recitative movement is used in and this is because we are able to get the emotions that are relayed and also the tonal variation are able to depict the support to the song and also the different voices that are used shows and proves that it is not dry but accompanied recitative. The three note theme is carried on by the strings. The third movement start to give form to the Cantata: instruments, recitative, instruments. The oboe introduction will trills and the bassoon are used as an accompaniment. The tenor comes as a solo but sings with the instruments. The constant movement of the instruments gives life. Jesus is maintaining life from Mary in the manger. The three note motif is repeated by the bass but the rhythm is jumpy. Is he the son of

Role government of each artical Research Paper Example | Topics and Well Written Essays - 2000 words - 1

Role government of each artical - Research Paper Example This will in turn allow the participants in the market to start putting the resources back to work in the sectors that they consider as most beneficial. This paper aims at considering the major roles of the government in the economic stability, with the inclusion of the government-run institutions. In the previous decades of lecturing countries that are developing on the ways to stimulate economic growth and emerge from economic crisis through strong government policies, some economists and U.S policymakers are discarding all the advice when subjected to the first test of major crisis. This is specifically true when taking advice regarding the accumulation of more debt (Shah, Rajiv, Zhijie, and Harini 85). For instance, the fiscal responsibility summit held by the U.S President Barack Obama last February disclosed that he knows the urgent requirement for fiscal responsibility. But the recent enactment of the Congress of the America Reinvestment and Recovery Act and the budget that was proposed by the government makes the objectives of a budget that is sustainable budget and it addresses the longer-term fiscal properties of the nation such as the entitlement liabilities, and even more elusive. Government deficits and government spending increase automatically when there are economic downturns that arise due to falling tax revenues and more demands on the social-safety-net provisions. Such spending can be helpful in attaining stability on the economy since it does not occur following legislative acts but automatically, and the money that is spent is usually needed most at the time of spending. Spending and borrowing for the purpose of economy stimulation by the use of legislative discretion is riskier since it is more difficult to wait for the right moment. The spending of the funds does not often take place until a long period has passed after the downturn occurs and has the ability of prolonging the downturn through crowding out

Giving Credit Essay Example for Free

Giving Credit Essay Abstract In this assignment we compare the lives of two men that created inventions that changed the world we live in. Throughout their careers and success’ they had hurdles to jump and become innovative thinkers to create futures in their challenging industry. Great men and thinkers can move mountains and open paths to new industries. Giving Credit Where Credit Is Due As entrepreneurs in a growing world of technology and change Andy Grove and Michael Dell pioneered the industry. They were innovators, visionaries and industry leaders in their fields. Andy Grove envisioned change to create faster and more productive ways to utilize the way we process information. As an immigrant he passed through the walls of terror in his home country of Hungry which was facing many issues of destruction that the Nazi’s enforced. He fled to the United States in the 1960’s to create a better life and future for himself. Andy established a small company that he called Intel which created an immense new future for the computer technology. His contributions awarded him the esteem acknowledgement as Time Magazine’s person of the year. He pursued his dreams as all inventors to create products and progress in the ever-changing need for new technology. Andy Grove created the chip and the microprocessor that allowed information to be processed much faster and smaller in size. Mr. Dell also an inventor at an early age set up a laboratory in his dorm room while attending college to create his personal computer. His success with the personal computer caught on quick because he used a direct theory to include his customer in the structure of his product. Michael Dell believed that understanding his customer’s needs and issues would enable Dell to create good competitive products. He believed that cutting the middlemen out of the sales market would allow him that advantage. With that direct approach Dell was able to endure many obstacles as well as the recession that created market crashes in the early 2000’s. (Krames, Jeffrey A. What the Best CEOs Know: 7 Exceptional Leaders and Their Lessons for Transforming Any Business) Both Andy Grove and Michael Dell endured much criticism from competitors  because of their business approach and products and business logic. Mr. Grove instilled paranoia in his corporate CEOs so that they could become fearful of being caught off guard. Mr. Dell used his customers to create standards that would give him the leading edge over his competitors. Both were accused of manipulating the industry and deceiving the product consumers. Intel was accused of creating an inferior product that put fear into the consumer to believe the chips that were being used would create disastrous failures to the products who endorsed his chip. Dell had a simple approach to create sales and marketing by lowering the cost of his product by cutting out the middlemen which made him more competitive and viable to sustain the economic downfalls. His competitors accused him of lowering his cost so as to drive the market sales away from the non profitable companies. Both Mr. Grove and Mr. Dell used simple methods to approach the needs and creativity of the growing markets. Andy Grove made sure that being too comfortable with the products that his company offered just wasn’t going to be enough. He looked ahead to keep creating new products that would soon be replaced by a growing need for better technology. Mr. Dell never doubted his approach to keep his overhead down and modeled his success on a customer based product. He believed creating his product for the customers rather than just offering his product without realizing their valuable input. Their similarities are based on what the customer and technology needs to sustain a profitable and sustainable future. They share innovative ideas in product development and customer based satisfaction. They differed by the way each approached the markets, Intel needed to create a superior product sooner and more often to keep up with changing technologies. Dell used the customer to create products based on the user need. Each approached their core values of business and what works for them to create value and sustainability. Dell had the market share by using the internet to generate sales through machine to machine based sales and conserved his manpower to keep the cost of his product low. Intel spent many hours of research and product development to create new and innovative ground breaking products that led the industry standards. Mr. Grove believed that being complacent would drive him out of the industry and eventually close the doors. He encouraged his CEOs to listen to his sales team to find out the needs of the customers Andy Grove referred to those individuals as â€Å"Cassandra’s† that would pass on valuable customer feedback. His pushes to create paranoia made Intel strong and create rather than become comfortable because Grove feared that waiting for the market to fail made Intel week. Mr. Dell believed that his customers gave him the strength to survive and customize his business to their needs. Dell never overstocked and always built their product to order. Dell realized that over burdening their inventory would devastate their ability to keep costs down. Dell learned from their mistakes like the Olympic brand which compromised the need of customers and their ability to utilize technology threatened the Dell line of products. Dell made efforts to speak to the public and hear what they had to say and put that information into their products. (Krames, Jeffrey A. What the Best CEOs Know: 7 Exceptional Leaders and Their Lessons for Transforming Any Business) Both of these pioneers created a wealth of knowledge and bridged the technology industry to what is today. Innovation is the foundation of the industry and creating new and exciting products that can be user friendly and affordable make profits. Both of these two companies are very profitable and determined to create intelligent and appealing industries. The changing markets constantly demand new and better technologies that will enable us to accelerate computer speeds and retain more memory without absorbing higher prices and engineering. References American Psychological Association. 2010). Publication manual of the American Psychological Association (6th ed.). Washington, DC: Author. (Krames, Jeffrey A. What the Best CEOs Know: 7 Exceptional Leaders and Their Lessons for Transforming Any Business)

Expression of Recombinant Green Fluorescent Protein (rGFP)

Expression of Recombinant Green Fluorescent Protein (rGFP) Expression and Purification of recombinant Green Fluorescent Protein (rGFP) from E. coli using Ni2+-Agarose Column Chromatography. Andrea Bustamante Janakikeerthika Darmarpandi Abstract Green Fluorescent Proteins are vital components of bioluminescence in marine animals. There unique ability to withstand and recover from harsh conditions and regain fluorescence was of great interest. The purpose of the following set of experiments was to express and purify a His6-Xpress epitope tagged recombinant form of Green Fluorescent Protein grown and harvested from E. coli. The desired protein is initially released into solution using the properties of freeze-quick thaw cycles that then help release the contents of the nucleus of neighboring bacteria following a chain reaction. It is then submitted through a Ni2+-agarose affinity chromatography column where the target protein was purified. The resulting wash and elution fractions where run through a Bradford assay, SDS-PAGE/Coomassie blue staining, and a Western blot to determine the molecular weight of the protein to be 32kDa. The overall specific activity was determined to be 433000 RFU/ mg of total protein with a resulting 20 percent purity. The results show that expression and purification of rGFP from bacterial cells was possible. Introduction Aequorea victoria is a jellyfish capable of producing a green fluorescent light when Ca2+ ions activate a photoprotein, known as aequorin, which excites Green Fluorescent Protein (GFP). Wild type GFP is a 27kDa, homodimer composed of 238 amino acid residues that absorbs light at an excitation wavelength of 395nm (blue light) and emits light at an emission wavelength of 510nm (green light). Aequorea victoria GFP has a distinctive three dimensional structure that encases a chromophore (formed by cyclization of Ser65-dehydrogenized Tyr-Gly67) and allows for stability under harsh conditions (Prasher, 229-230.) . This structure allows for regaining of fluorescence even after the protein has been denatured upon removal of the denaturant. Therefore, GFP’s are extremely stable to changes in pH, temperature, oxidation and reduction, and chemical reagents (Pan, Pickett, and Rippel 225.) Poly-histidine tags involve addition of a series of histidine residues to the N or C terminus of a protein of interest. Poly-histidine tags are affinity tags that serve to facilitate protein purification by exploiting the positively charged histidine residue’s affinity for negatively charged columns. This series of experiments involved a six repeat histidine codon contained within a DNA plasmid which resulted in a recombinant Green Fluorescent Protein that contained a six residue histidine tag located at the N-terminus. The His ­6 tagged recombinant Green Fluorescent Protein was then subjected to Ni2+-agarose column affinity chromatography. Ni2+-agarose affinity chromatography allows for the purification of poly-histidine tagged proteins due to the selectivity and affinity of the Ni2+-agarose matrix for His6 tagged proteins. rGFP binds the column due to the interactions between the His6 tagged proteins in the mobile phase with the metal Ni2+ ions immobilized within the matrix in the stationary phase. The Ni2+ ions contained within the matrix are capable of binding electron rich molecules including histidine residues and allowing most other molecules to pass unbound. This results in the binding of the desired protein to the column and the purging of most undesired proteins and contaminants from the column into wash fractions (Ninfa, et al. 100-101.) The column was then subjected to imidazole, which competes with rGFP for Ni2+ ion attachment, and this allows for the elution of the target protein. Due to its unique properties, isolation of GFP was of great interest and expression and purification were the main focus of the following series of experiments. A suitable way to accomplish this was devised using the combination of poly-histidine tagging and affinity chromatography. The purpose of this experiment was to express and purify a six-Histidine tagged recombinant form of Green Fluorescent Protein from E. coli through the use of Ni2+-agarose affinity chromatography. After expression and purification, a Bradford assay was performed to estimate total protein amount. This was followed by SDS-PAGE/Coomassie blue staining to determine purity and molecular weight. The confirmation of the presence of rGFP was done using the Western Blot. Materials and Methods Growth of G strain In a test tube, 10ml of liquid LB growth media containing 100ug/ml Amp and 25ug/ml Cam was inoculated with a single bacterial colony of strain G (BL21(DE3)uv>) and was allowed to grow overnight at 37 °C. The culture was shaken until saturated. In a flask, 500ml of liquid LB media (pre-warmed to 30 °C) was inoculated with about 4 ml of the saturated overnight culture (or until the 500ml culture reached an OD600 reading of 0.1) and allowed to grow at 37 °C until the OD600 reading reached 0.5. At approximately OD600 ~0.5, or time zero, 1ml of the culture was harvested into a 1.5ml centrifuge tube and pelleted. The supernatant was discarded and the â€Å"G0† pellet stored at -20 °C for later use. The remaining culture was induced with 1mM IPTG and allowed to grow. After 3 hours, 1ml of the culture was harvested into a 1.5ml centrifuge tube and pelleted. The supernatant was discarded and the â€Å"G3† pellet stored at -20 °C for later use. An additional 15ml of the IPTG induced culture was harvested into a 15ml centrifuge tube and pelleted. The supernatant was discarded and the â€Å"G3-15ml† was stored at -20 °C. Preparation of rGFP Crude Extract Immediately after removal of the â€Å"G3-15ml† pellet from freezer, breaking buffer [10mM Tris, pH 8.0; 150mM NaCl] was added into the centrifuge tube. The breaking buffer was pipetted up and down (being careful not to introduce air) until pellet had thawed and homogeneity was reached. The solution was transferred into a 1.5ml centrifuge tube, vortexed for 5 minutes, labeled and placed in 37 °C water bath for 10minutes after which the centrifuge tube was transferred to a rotating platform shaker in a dry air 37 °C incubator for 20 minutes. After lysis, the mixture was centrifuged at 14000xg, 4 °C, for 10 minutes. In a dark room in the presence of a hand held UV light, the fluorescence of the pellet and supernatant where observed the recorded. The supernatant was then decanted and care was taken not to get the pellet back into the supernatant as centrifugation would be required if this did occur. This supernatant was the GCE (rGFP crude extract) Preparation of Ni2+-agarose Column In a 3ml plastic syringe, enough glass wool was placed into the well to cover up to the 1/4 ml marking. The syringe was secured onto a ring stand and placed perpendicular to the ground. About 100ul of breaking buffer was pipetted into the top of a closed luer-lock and allowed to overflow. 1ml of buffer was then pipetted into the syringe column and the luer-lock was immediately screwed onto the syringe. An additional 2ml of breaking buffer was added to the column and several drops of buffer were allowed to flow out. The luer-lock was then returned to the closed position. A total of 500ul of breaking buffer was added to the column and then 1ml of a 0.5ml bed volume Ni2+-agarose slurry was added to the column. The luer-lock was opened and agarose matrix was allowed to â€Å"gravity pack.† The column was pre-equilibrated with 5ml of breaking buffer and then the luer-lock was returned to the closed position. Ni2+-NTA Chromatography Separation Procedures 100ul of GCE was transferred into a centrifuge tube, labeled, and set aside. Breaking buffer was added to remaining GCE if content was less than 1ml. GCE was slowly applied to the Ni2+-agarose column and allowed about 5-10 minutes for protein to bind to column. The luer-lock was opened and 0.5ml of effluent was collected into 1.5ml centrifuge tube and labeled W1. This was repeated with the subsequent effluent labeled W2.The column was then observed under an ultraviolet light and fluorescence recorded. The column was then washed with 4ml of buffer in 0.5ml increments. The effluent was collected and labeled W3 to W10. The column was then washed again with a total of 5ml of breaking buffer. This effluent was discarded. A total of 5ml of elution buffer containing 10mM Tris, pH 8.0; 150mM NaCl, 300mM imidazole was added to the column in 0.5ml increments. The eluents were collected and labeled E1-E10.The column was then observed under a UV light and the fluorescence recorded. The W1-W6 and E1-E6 fractions were also observed under UV light and their fluorescence rec orded qualitatively. Determining Total Protein Amount A standard curve was created using six different samples of Bovine Serum Albumin (1mg/ml) of known amount. The amounts of BSA used all had a final volume of 50ul and included 0ug, 3ug, 5ug, 10ug, and 20ug total proteins. A total of 1ml of Bradford reagent was added to each, vortexed, and allowed to incubate for 10 minutes. The results where read using 200ul in a microtiter dish and read using a microplate reader set to 595nm. The results where plotted on a graph as absorbance (595nm) vs. BSA (ug) and a best fit line was drawn. The Bradford assay was then performed once on the W1-W6 and E1-E6 samples. Any samples whose absorbance fell outside the standard curve were repeated less sample in the assay. Once all samples fell within the standard curve, the Bradford assay was repeated two more times for each sample. The total protein amount was then extrapolated from the standard curve using the absorbance values. Estimating Purity and Molecular Weight The SDS-PAGE was prepared using a 12 percent resolving gel that was poured between the Bio-Rad glass plate â€Å"sandwich† and allowed to polymerize. A 5 percent stacking gel was prepared and added on top of the resolving gel, a comb was inserted, and the gel was allowed to polymerize. Once that polymerized, the combs were removed and the electrophoresis tank was set up. 15ul of G0, G3, GCE, W3, W4, E2, and E3 samples were added to the SDS-PAGE along with a standard molecular weight ladder. The samples were electrophoresed at 200volts for 45 minutes. The gel was then stained using Coomassie blue dye and the stain removed. Confirmation of rGFP 2-ÃŽ ²-mercaptoethanol was added to the centrifuge tubes containing the G0, G3, GCE, W3, W4, E2, and E3 samples and were loaded along with a molecular weight ladder and electrophoresed as described above. The stacker was removed and the resulting gel set up for transfer onto a nitrocellulose membrane for Western Blot analysis. The overall setup required a â€Å"building up† of components with the positive electrode base on the bottom, followed by filter paper soaked in transfer buffer, nitrocellulose paper above that, the SDS/PAGE layer, another layer of filter paper soaked in transfer buffer, Western blot solution was poured over all the components, and finally the negative electrode lid was locked into position. To ensure transfer, the nitrocellulose gel was stained using Ponceau S and allowed to incubate for two minutes on a rocker and then destained using ddH2O. The membrane was then blocked using 5% non-fat dry milk/TBS solution and incubated for 30 minutes on a rocking p latform. This was then and washed three times with 0.05%Tween 20/TBS with 5 minutes of incubation between each wash. It was then probed with mouse IgG anti-Xpress epitope MAb solution and allowed to incubate for 45 minutes. The 0.05%Tween 20/TBS wash was repeated in triplicate. A secondary probe using sheep IgG anti-mouse IgG conjugated horseradish peroxidase polyclonal anti-serum solution was performed as above and then washed in triplicate. The nitrocellulose gel was developed using TMB until desired intensity was reached and development was stopped with water and results recorded immediately. Results The expression of the target protein was doubly repressed in the G0 (uninduced) sample of E. coli. First, the Lac repressor protein binds to the lac operator and prevents transcription by T7 RNA polymerase (Garrett and Grisham 915-916). Second, T7 RNA was repressed by lysozyme protein that binds to T7 RNA polymerase and inhibits transcription. Expression of rGFP in the G3 (3 hour post induction) sample was made possible through the use of IPTG (Garrett and Grisham 914.) The purpose of IPTG was to repress the Lac repressor which resulted in T7 RNA polymerase being able to transcribe DNA downstream of the T7 promoter and expression of His6-Xpress-GFPuv, resulting in the fluorescent capable recombinant Green Fluorescent Protein. (Figure 1) This resulting recombinant GFP is a 279 amino acid protein. rGFP has a six Histidine tag at its N terminus between amino acids 5 and 10, an Xpress epitope between amino acids 24 and 31, Green Fluorescent Protein between amino acids 39 and 277, and a 3 amino acid end tag between amino acids 277 and 279. The chromophore is found between amino acids 103 and 105 in the DNA sequence. (Figure 2) Results of Ni2+-agarose affinity chromatography and Bradford assay indicated that the E3 (elution 3) sample contained the most rGFP activity with approximately 18,600 RFU (relative fluorescent units) and an estimate 43ug of total protein. The specific activity calculated for the sample was 433000 RFU/ mg of total protein. (Figure 3) The SDS-PAGE/Coomassie staining gave an estimate molecular weight for rGFP of 32kDa based on a total traveled distance of 2.3cm along the SDS/PAGE. The overall purity of the band was approximately 20 percent. The higher molecular weight band was most likely contaminants at about 45kDa and the lower molecular weight band was possibly a result of the degradation of the c-terminus at 27kDa. (Figure 4) Western Blot indicated prominent bands in the E3, E2, GCE, and G3 lanes. Lanes W4 and W3 showed very light bands and lane G0 shows an absence of bands. All visible bands appear at about 32 kDa and therefore confirm the presence of rGFP. (Figure 5) Conclusion The successful expression and purification of recombinant Green Fluorescent Protein is significant in the scientific community due to the possible uses for it in the future. Green Fluorescent Protein is significant because it provides an inexpensive and relatively easy method of detection. The possibility for real time detection means result could be obtained in real time. Future experiments will focus on linking rGFP to proteins during transcription and translation. This would result in a desired protein with a GFP tag whose fluorescence can then be used for identification. This should result in the ability to locate a target protein using the fluorescence of rGFP. Future applications of GFP could include incorporation into the genetic code of small mammals. These could encode fluorescent neurons which in turn could help further research in areas such as nerve tissue regeneration or other advances in neurobiology. Its unique properties of endurance could be exploited to understand how it can endure harsh environments and still regain functionality after remediation. This would have significant applications in molecular and cellular biology in understanding cellular degeneration and how help patients with diseases involving cellular degeneration. Bibliography Pan, Jing, Elizabeth Pickett, and Scott Rippel. Biochemistry Laboratory Lecture Notes. Dallas: UTD copy center, 2013. 225-289. Print. Pan, Jing, Elizabeth Pickett, and Scott Rippel. Biochemistry Laboratory Manual. Dallas: UTD copy center, 2013. 38-77. Print. Prasher, Douglas C., Virginia K. Eckenrode, et al. Primary Structure of the Aequorea victoria green-fluorescent protein. Gene. 111. (1992): 229-233. Print. Garrett, R., and Charles M. Grisham. Biochemistry. 4th ed. Belmont, CA: Brooks/Cole, Cengage Learning, 2010. Print. Ninfa, Alexander J., and David P. Ballou. Fundamental laboratory approaches for biochemistry and biotechnology. Bethesda, Md.: Fitzgerald Science Press, 1998. 89-107. Print.