Project proposal for summer 2005

Aaron Holman

How can a complex behavior, such as addiction, be affected by a single gene? To answer this question a combination of behavioral studies and genetic profiling must be used. Once a gene has been identified as having a link to a behavior, in this case ethanol consumption, researchers need to determine what effect it has on said behavior. The best way to do this is to cause the neural cells to over express the gene and then observe the change in behavior. One challenged faced by researchers is that the most commonly used vector for injecting the gene into cells, AAV, only infects dividing cells. In order to infect neurons, the gene of interest must be transferred to a lentivirus vector, which readily infects non dividing cells. My goal for the first summer is to accurately transfer the sgk gene into the lentivirus vector so that the effects of this gene on drinking behavior in mice can be observed.

In order to get enough of the vector to infect the desired tissue, large amounts of plasmid containing the gene are needed. The plasmid will be cloned through transformation of competent cells and then extracted and purified. In order to determine the quality and validity of the plasmid, it will be sequenced and matched against the expected code. The purified plasmid stock will be sent to a separate lab to be placed into the lentivirus.

If procedures are followed correctly, the sgk gene should remain intact through the entire process. Mishandling of the samples can lead to shearing of the DNA and contamination may also hinder results.