Abstract:

In order to study the function of flavocytochrome C (flac) in the producing-hydrogen metabolism, the authors used CODEHOP to design the degenerate primers of flavocytochrome C, chose one pair of degenerate primers named FlacJ, then used Enterobacter cloacae NRRL B-414 genome DNA as template to make degenerate PCR, got about 1740 bp PCR product, they were transformed into the E.coli DH5α through being linked with pMD18-T vector and sequenced after filtration. The fragment is 1740 bp long, the G+C content is 58.51%, the A+T content is 41.49%, and its NCBI accepted number is GU565952. Similarity alignment showed that the 1740 bp products were very similar to the flavocytochrome C genes, and shared 95% identity to the large subunit of flavocytochrome C from Enterobacter cloacae subsp. cloacae ATCC 13047, and only had the difference of twenty-eight amino acids. Cloning this fragment would not only enrich the gene resources of flavocytochrome C genes, but also give the scientific warrant for the metabolic engineering research. And it was proved once again that using CODEHOP to design degenerate primers for cloning homologous family genes has a high success rate.