Construction And Detection Of CG31007Bait Plasmid

The body colour of spontaneous mutations in Drosophila melanogaster strains black streak(BSR)appear complex characters:deep black, brown and pale yellow. To investigate the molecular mechanisms of complex characters, by Blastp analysis, it showed that the corresponding protein CG31007-PA possessed a conserved domain.We predict that the CG31007protein is related to the function of cytochrome C oxidase. Cytochrome C can oxidize catecholamines into melanin by H2O2in mitochondria, suggesting that CG31007mutations may cause the formation of melanin, thus affecting the Drosophila’s body color. The trial designed primers based on the CG31007ORF sequence, the target gene was obtained by PCR. Then by constructing a high expression vector that was transformed into E. coli, a lot of the target gene was gotten. By the yeast two-hybrid system, pGBKT-CG31007was constructed, then it was transferred into yeast AH109, and conducted toxicity and self-activity detection, it indicated that the recombinant plasmid for yeast growth is non-toxic, and can not be self-activated. This recombinant plasmid can be used as the "bait" for yeast two-hybrid system in the later trial, to screen the protein that can instruct with CG31007in the corresponding Drosophila cDNA library. By the extraction of total RNA, reverse transcribed into cDNA, to lay the foundation of the construction of Drosophila cDNA library in yeast for the future.