Purify GFP From Aequorea Victoria

Purify GFP From Aequorea Victoria Methods and Results: GFP was cloned into E. coli strain JM109 and expressed under optimal conditions in Luria broth agar containing ampicillin and IPTG for induction. Protein was extracted by lysis using bead milling technique and fluorescence of protein measured in a fluorimeter, concentration of both pure and crude proteins were obtained with Bradford (1976) method. Purity of GFP was further confirmed by SDS-PAGE stained with coomassie blue. Conclusion: Specific activity(RFUmg-1) of pure protein increased compared to crude representing increase in purity, with a substantial yield of 82%. Significance and Impact of Study: This study proved Ion exchange chromatography as a reliable technique for GFP purification and high percentage recovery for use as a reporter gene in molecular biology studies. Keywords: GFP, purity, Ion exchange chromatography, Specific activity, fluorescence. INTRODUCTION The jelly fish Aequorea victoria, emits a bluish light from the margin of its umbrella (Inuoye, and Tsuji 1994). The light is produced by the bioluminescent jellyfish when calcium binds to the photoprotein aequorin. Although activation of aequorin in vitro or in heterologous cells produces blue light, the jelly fish produces green light. This light is the result of a second protein in A. victoria that derives its excitation energy from aequorin, the green fluorescent protein (Chalfie et al., 1994). Green fluorescent protein (GFP) is a protein of 238 amino acid residues. It is a highly stable protein possessing a tightly packed ÃŽ ²ÃƒÅ'†°ÃƒÅ'†°ÃƒÅ'†°ÃƒÅ'†°can tertiary structure that is resistant to many biological denaturants, most proteases, pH (5-12), temperature (Tm=78  °C), and chaotropic salts (McRae et al., 2005; Zhuang, et al., 2008). Purified GFP absorbs blue light (maximally at 395nm with a minor peak at 470nm) and emits green light (peak emission at 509nm with a shoulder of 540nm). This fluorescence is stable and virtually no photobleaching is observed (Chalfie et al., 1994). The stable and intense fluorescence of GFP without any cofactors in many different organisms makes is ideal for molecular biology applications such as markers for gene expression analysis of molecular interactions and also as biological information storage devices and optical biosensors in areas of Nanotechnology (McRae et al., 2005). Current purification procedures, specific for GFP include multiple phase high performance liquid chromatography (HPLC), chromatofocusing on a pH gradient, metal ion precipitation and organic extraction. Most of these methods are either expensive, time consuming or give low yields with

Homosexuals in the United States Military Essay

Homosexuals and U. S military service new laws and regulations which came into effect in 1993 reflected a compromise in policy. This settlement, referred to as â€Å"don’t ask, don’t tell,† states that the existence in the armed forces of persons who reveals a tendency or plan to take on homosexual acts would produce an intolerable hazard to the high standards of morale, high-quality order and discipline, and unit solidity which are the core of military capacity. Service members are not to be asked about nor allowed to talk about their homosexuality. This negotiation in spite of the matter has remained politically controversial. Previous to the 1993 compromise, the figure of individual’s releases for homosexuality was by and large declining. From the time, the number of discharges for homosexual conduct has generally amplified until recent times. In the wake of the new 1993 laws and regulations, constitutional challenges to the former and current military policies regarding homosexuals followed. In the case of Bowers vs. Hardwick, the U. S Supreme Court Ruling said that there is no right to engage in consensual homosexual sodomy. In this case, the courts generally said that military men may be lawfully discharge for explicit homosexual conduct. Nevertheless, the legal picture was convoluted by the Court’s 2003 decision in Lawrence v. Texas which ruled against Bowers by declaring unconstitutional a Texas law that prohibited sexual acts between same sex couples. Moreover, disturbed legal questions lingered as to whether a release based exclusively on a statement that a service member is homosexual disobeys constitutional limits. For the time being, efforts to allow individuals of the same sex to marry legally materialize implausible to affect the Department of Defense (DOD) policy close to term, because such individuals are barred from serving in the military, even though court challenges are possible. For the duration of the 1992 presidential campaign, Bill Clinton promised to remove the ban on homosexuals in the U.  S armed services. Once in office, he met with massive resistance from the U. S military and its congressional allies, and by summer of 1993, the original policy proposal was dead. Instead, Congress enacted the â€Å"Don’t Ask, Don’t Tell, Don’t Pursue† policy: gays and lesbians can now serve in the military, but they are obliged to keep their sexual preference private. Challengers of the open integration of gays and lesbians have discarded many of standard justifications for excluding homosexuals from military service. For example, the Pentagon and its cronies no longer disagree that gays and lesbians are security risks because of the threat of blackmail. As a case in point, even though both the former chairman of the Joint Chiefs of Staff General Colin Powell and the famous military sociologist Charles Moskos contest the open integration of homosexuals in the military, they recognize that gays and lesbians are valuable soldiers. Discharge measures against homosexuals are packed with statements of many of these individuals’ excellent records, reliability, and commitment to their jobs. The matter is not whether gays and lesbians are good quality soldiers as individuals, but instead, the consequence of these individuals on the group. Opponents of removing all restrictions on homosexuals’ service argue that open incorporation of gays and lesbians would obstruct the development of primary group cohesion, which they say is significant to military efficiency. During the 1993 congressional trials on homosexuality in the military, both Senate and House testimony paid attention on the issue of unit cohesion. For instance, then Senator Sam Nuun, chairman of the Senate Armed Services Committee in July 1993 asked each of the 6 Joint Chiefs of Staff to discuss unit cohesion and its importance in developing combat capability. Army Chief of Staff General Gordon Sullivan answered him by saying that cohesion is developed by uniformity, by devotion to a common sense of values and behavior. The introduction into many small units of person whose open orientation and self-definition is completely opposed to the rest of the group will cause tension and disruption (Herek 1987). Senior US military officers concerned that the open integration of homosexuals would get in the way of the development of cohesion within small groups are not trusted or respected as expressed by Powell and Admiral David Jeremiah, and they added that in atmosphere of doubts, orders may not be carried out and everyday friendly gestures that encourage companionship- everyday childlike horseplay and rough-housing, a pat on the back or arm around the shoulder- become suspect, provoke fear or loathing, and annihilate group cohesion (Nowak, 1993)). Powell added that in order to win wars and battles the army needs to make cohesive groups of warriors who will bond so strongly that they are ready to go into battle and give their lives if indispensable and it is intolerable to allow anything to upset that feeling of cohesion inside the force. The disagreement about unit cohesion is based on two propositions: the first one is that primary group cohesion increases military effectiveness and second, open gay and lesbian personnel would disturb cohesion and thus military performance. According to Kier (1998) these propositions are wrong and she said that such statements do not reflect what social science research and experience have demonstrated about the relationship between cohesion and performance and the consequence of putting together previously excluded groups on primary group cohesion. Investigations of Homosexual Conduct Even if broad investigations of homosexual behavior are the exemption rather than the rule, there are noteworthy numbers of cases in which such investigations have been conducted. Based on the cases reviewed by Gosling (1993), he concluded that the immense majority of investigations that have happened have been correctly instigated, that is, an investigation has been made only after the commander had determined that there was convincing information that the member had engaged in homosexual conduct. Also, based on his findings, he was able to find out that a lot of the criticisms made about inappropriate initiation of investigations mirror a misinterpretation of the Department’s policy. In practice, plausible information has sometimes been provided to commanders in ways that service members might not have been anticipated to occur, or has been based on communications or performance that the partners, roommates, or unconnected third parties have sometimes come forward on their own to account information or proof of homosexual conduct to commanders next to the wishes of the service member in question (Nowak 1993). Photographs or in black and white communications that verifies homosexual conduct has sometimes been showed to civilians who then brought this proof to the attention of a commander, with no question having been conducted by the commander. Plausible information has also been incidentally discovered in the course of proper, entirely unrelated criminal or disciplinary investigations for a commander to initiate an investigation when information has been reported in any of these circumstances, granted that the information received is credible (Suraci, 1992). Indeed, because federal law requires that those who take on in homosexual conduct must be discharged from the military; commanders are compelled to investigate whenever they receive credible evidence of homosexual conduct. In addition, many academic institutions have ratified rules that defended homosexuals from prejudice on campus. Accordingly, colleges, universities, and even high schools have required barring military recruiters from their campuses or otherwise eliminating Reserve Officer Training Corps Programs on campus because of the DOD rule on homosexuals in the military. Simultaneously, legislation has been ratified that bars giving federal funds to campuses that obstruct entrance for military recruiters (Suraci 1992). On March 6, 2006, the Supreme Court upturned a federal appeals court verdict in Rumsfeld v. Forum for Academic and Institutional Rights (FAIR), and endorsed the constitutionality of the Solomon Amendment, which forbids certain federal funding to higher educational institutions that refute admittance by military recruiters to their students equal to that provided to other employers. Deviance Present among sociological conceptions of deviance is an approach that concerns itself not as much of with the characteristics of the person or persons said to have despoiled a social rule than with the character of the responses of other persons to these characteristics and events. This approach, occasionally called the â€Å"labeling† approach to deviance, observes the deviant as a social creation, the result of contact sequences between labelers and labeled (Becker, 1964). The questions that are lifted by this approach thus concern the behaviors that are tagged as deviant, what the methods are by which the labels are effectively applied or avoided, and what the results of such procedures are for both labelers and labeled. Nevertheless, regardless of the highlight laid on deviance as a creation of interaction, in practice most consideration has been paid to the labeler’s role in this process (Simon 1987). The method of developing deviance appears all collective response and no deviant stimulus. This is possibly an overreaction to an overreaction. For example, one theory is spelled out as to why some people break rules and some do not in terms of what is called as commitment and for other labeling theorists, characteristics of the deviant himself are not completely unrelated (Simon 1987). The point is well taken, but for example, in some of Goffman’s writings (1961) on mental patients his employment of the concept career contingencies materializes to treat the mental patient as a pawn, subject to the vagaries of all sorts of contextual demands. As a result of the above, labeling theory also has been seen as taking the side of the underdog- that the deviant is seen as a victim of the fairly subjective measures of control agencies. He is more sinned against than sinning, as it is a matter of chance, or racial or socioeconomic factors, rather than any behavior on his part, that chooses whether he is cast as deviant. Homosexuals in the Army in Other Countries Homosexuals in the military is not just an issue faced by the United States, there are many more countries out there that have the same situation as the US. The number of countries that permit gay and lesbian soldiers to serve in the armed forces is growing and it is increasingly becoming more important to know whether official decisions with regards to the inclusion of homosexual service members in the military lead t changes in organizational performance (West 1965). Even though most members of NATO plus some nations has already permitted gay and lesbian soldiers to serve, there has still a very few empirical analysis of whether the decision to remove gay ban influences the ability of armed forces to pursue their missions. This topic has been addressed by some theoretical studies but there has been no in-depth empirical analysis on the consequences of removing gay bans (Sudnow 1965). In Canada for example, there were a handful of careful studies immediately after Canada’s 1992 decision of abolishing restrictions on gay and lesbian soldiers (Belkin, 2001). Although that was the case, long term impact of the new policy could not be known in those early studies and even the best qualitative research is only based on a few sources (West 1965). American officer, Lt Gen Calvin Waller, affirmed in 1993 that since Canada had not been caught up in armed conflict since the ban was lifted, he explained that Canada’s justification for considering the proof that has mounted up for up to eight years since the ban was removed is that senior Canadian officials foresee that altering the policy might compromise military effectiveness (Millet 1999). For this reason, the Canadian incident gives an opportunity to assess the effect of the policy change in opposition to early forecasts by senior military leaders. After discussing the historical development of homosexual personnel policy in Canada, the authors examined whether Canada’s decision to eliminate restrictions on gay and lesbian soldiers influences military efficiency (Brumett, 1981). The paper’s findings, based on a review of primary and secondary sources, as well as interviews with 29 military personnel and experts from the academic, non-governmental, and policy communities, is that Canada’s decision to lift its gay ban had no effect on military performance, eagerness, unity, or morale (Millet 1999).

The Modern Alterations of Native American Life

Modern Native American life has changed drastically from that of what it used to be. One can only imagine how incredibly hard life would have been in the late 1800s without the help of modern technology. In James Welch's book, Fool's Crow, one not only learns what the Western United States was like in the late 1800s, but can also grasp the ways of early Native Americans. Fools Crow, the main character in the book, portrays exactly what the average Indian boy of his day was like. He had strong family values, faithful religious beliefs, and knew his direction in life. Unfortunately this is not so true for most Native Americans of today. Many current Indians are suffering from poor family structure, lack of education, and loss of religious beliefs. Family structure was one of the most important aspects of life for Native Americans of the 1800s. Back then there was almost no difference between an Indian man or woman's fifth cousins and his or hers sister. Both were treated the same, and both were respected immensely. But now the family structure of Native Americans has diminished. The communication between the majority of parents to children is very slim, if any, and many young Native Americans don't even have contact with there grandparents and other extended family. In Fools Crow's generation it was very common to often have ceremonies where all members of a family were expected to attend, whether they were a grandparent or a brother. This failure of family structure in modern Indians has caused many individuals to lack security and subsidity. Education is a problem among Native Americans as well. It used to be that when a young Native American turned a certain age it was his or her duty to learn the ways of survival. For boys it was hunting and learning to provide for his future family. For the women it was learning various duties such as cooking, tanning hides, and caring for the children. Indians used to educate themselves in order to survive as a team. Each person learned to do his or her specific duty in order to prosper individually, and as a tribe or band. But now education is substantially different. It still focuses on the essentials for survival and the need for one to prosper as an individual, but it lacks what white people are learning. Teachers on reservations are often the product of poor education themselves, and have nothing to offer young Native Americans. One of the most prominent problems, however, is that reservations can't afford to hire quality teachers to educate their children. Thus, there continues to be uneducated Native Americans, which, in turn, causes this misfortunate cataclysm to continue. Another, and most important, aspect for a specific group of people to thrive, is religion. Ever since the first civilized human lived, there was religion. For most people, religion is the glue that holds their life together. It helps to provide purpose and meaning to all life. Native Americans of the 1800s, and before then, held very strongly to their beliefs. Religion was in everything they did. Whether it was planting crops or going on a big hunt, religion was the key to it. But now the Native American's religious beliefs have faded dramatically. Most of today's Indian youths no longer believe or practice the religion of their ancestors. Without religion, modern Native Americans find themselves insecure in what their purpose in life is and they often lack moral responsibility as well. The key to possibly fixing this problem is to educate young Native Americans about what their people used to believe in, and what role it played back then. With some good leadership skills and a lot of enlightening, maybe the old Indian religion will be fully restored to what it once was. In conclusion, Native American culture is rapidly being destroyed. Modern Native Americans have lost what it takes to keep a culture alive; religion, structure, and education. If Native Americans are once again going to flourish as a ethnic society, people need to do whatever they can to help build back the essential parts of Native American life.

Business Management Essays - 1108 Words

Business Management The world of business has undergone radical and dramatic changes in the last decade changes that present extraordinary challenges for the contemporary manager. A manager is an organizational member who is responsible for planning, organizing, leading, and controlling the activities of the organization so that the goals can be achieved. According to a widely referenced study by Henry Mintzberg, managers serve three primary roles: interpersonal, informational, and decision-making. Management is process of administrating and coordinating resources effectively and efficiently in an effort to achieve the goals of the organization. The concept of management within an organization typically occurs in an†¦show more content†¦Controlling is monitoring the performance of the organization, identifying deviations between planned and actual results, and taking corrective action when necessary. With all these four functions that are involved in the process of management, if all are followed correctly the organization will be properly ran and will have few complications. In every organization there are managers, and every manager is classed on different levels in terms of the skills they need and the activities they are involved in. Managers exist at various levels in the organization hierarchy. A small organization may have one layer of management, where as a large organization may have several. In most organizations there are three level of managers. The three level of managers are, first line manager, middle manager, and finally the top-level manager. Managers at different job levels have different job responsibilities and therefor require different skills. First-line managers supervise the individuals who are directly responsible for producing the organization’s product or delivering its service. 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