Advisor(s) - Committee Chair
Claire Rinehart, Valgene Dunham, Frank Toman
Department of Biology
Master of Science
A plasmid (pGEM-SD) was constructed in which the Shine-Dalgarno sequence had been deleted from pGEM-7zf(+) plasmid to generate a vector for regulated expression of antisense RNA. The binding of antisense RNA to mRNA provides a potent mechanism by which specific transcripts can be translationally inactivated. Although part of the lac operator sequence was deleted in pGEM-SD plasmid, it was proven that mRNA still can be induced under the control of lac promoter. Recombinant plasmids were generated by ligating bacterial genornic DNA into pGEM-SD plasmid, but the orientation of the inserted gene with respect to the lac promoter has not been determined.
These preliminary results show that gene expression can be turned on and off via the pGEM-SD vector. Future experiments will screen antisense recombinant clones for those which would inhibit specific gene products by plating on different selective indicator media both with and without the inducer IPTG. This will allow investigation of the biological functions associated with genomic DNA sequences by means of gene mapping with antisense RNA.
Biology | Life Sciences
Wu, Wen-Jun, "Creation of a Vector for Regulated Expression of Antisense RNA in Escherichia Coli" (1992). Masters Theses & Specialist Projects. Paper 3003.