SampleProposal2 - S u ic id a l G e n e E x p r e s s io n...

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Suicidal Gene Expression in Isolate Bacteria for Oxidization of Hydrocarbons in Crude Oil [Authors names removed] ABSTRACT One of the more controversial issues concerning hydrocarbon-oxidizing bacteria, which can be used in the bioremediation of contaminated water, is whether the disruption caused by introducing non-indigenous organisms into an existing ecosystem in the environment will be more harmful than the original contamination. This project will investigate the properties of the hok/sok gene system found in the R100 plasmid of E. coli and assess whether the gene can be incorporated into strain TD3, a hydrocarbon-oxidizing bacteria. If successful, the hok gene will be expressed after the strain TD3 bacteria oxidizes all the contaminants in the water, leading to a “suicide” of the bacteria. This way, the bacterium will undergo programmed cell death after it decontaminates the water, leaving no foreign microbes behind to disrupt the original ecosystem. This use of naturally occurring “killer genes” in genetically engineered microorganisms could be a resolution to the ethical issues of using genetically altered organisms for bioremedial purposes. OBJECTIVE One of the more controversial issues concerning hydrocarbon-oxidizing bacteria, which can be used in the bioremediation of contaminated water, is whether introducing non-indigenous organisms to an environment will disrupt the existing ecosystem. This has inhibited the application of research involving genetically engineered microorganisms, a potential method for degrading crude oil spills. The proposed solution is to introduce an agent into a bacteria’s genome that would cause cell death by “suicide” following the complete consumption of hydrocarbons in the oil. This would not only allow for the safe use of bacterium for contaminated waters, but pave the way for a safe and accepted use of genetically altered microbes for bioremedial purposes. INTRODUCTION Bioremediation focuses on using the natural processes of certain microorganisms to remove harmful chemical substances from the environment. Some types of bacteria are actually able to degrade matter such as hydrocarbons from crude oil, because they use these compounds as carbon sources. The decomposition breaks down potential toxins into CO 2 and other less damaging compounds. 1 One species of bacteria, known as Desulfothermus naphthae (TD3 strain), is naturally occurring and has the ability to oxidize hydrocarbons found in crude oil. 2 In previous experiments, the bacteria strain was capable of oxidizing saturated hydrocarbons ranging in composition from six to sixteen carbon atoms, which covers a majority of petroleum compounds (usually C 5 to C 18 in length). 3 This process of breaking down such substances is known as biodegradation, a form of bioremediation, and results from other experiments show that genetically modifying such microbes by bioaugmentation increases the rate of hydrocarbon oxidation.
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