Identification of the role that the 3'UTR plays in regulation of Hdc expression in Drosophila
Genomic un-translated regions (UTR’s) do not code for proteins and until recently were believed to lack functional purpose in the genome. Current studies indicate, however, that 3’UTR’s play a role in the regulation of gene expression. Last summer our lab conducted research on this topic, fusing the putative promoter region of the Hdc gene of Drosophila melanogaster to enhanced Green Fluorescent Protein (eGFP) to create a new transgene, pHdc-eGFP. After integration of the transgene into the Drosophila genome, an incomplete pattern of GFP expression compared to Hdc’s pattern of expression was observed. This result indicates that there is another genomic region that is required for complete expression. Therefore, we have been working to construct a new transgene that incorporates the 3’UTR of Hdc into the pHdc-eGFP transgene. Upon completion, this transgene can be used to analyze the effects of the 3’UTR on gene expression. Using PCR, we amplified a 1.13kb NheI-SpeI GFP fragment from the pHdc-eGFP transgene and TA cloned the fragment into the pGEM-T Easy Vector. We performed the same steps to obtain the 0.9kb 3’UTR of Hdc from the pCasper3-gHdc plasmid. Next, we PCR amplified two fragments from the NheI-SpeI GFP + pGEM-T Easy plasmid, one reading from NheI to the first termination codon of GFP and the other reading from the second termination codon of GFP to SpeI. Both fragments have been TA cloned and we are awaiting the results of sequence analysis before moving on to the next step towards completion of the new pHdc-eGFP-3’UTR transgene.
Faculty Mentor: Martin Burg, Biomedical Sciences
Embriette presented at the 51st Annual Drosophilia Research Conference April 7-11, 2010 in Washington DC.
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