Abstract:

In order to study the function of formate hydrogen lyase repressor (hycA) and large subunit of hydrogenase 3 (HyA3) in the producing-hydrogen metabolism, the authors used CODEHOP to design the degenerate primers of hycA and HyA3L, and chose two pairs of degenerate primers named HyA3J and HycAJ respectively, then used Enterobacter cloacae NRRL B-414 genome DNA as template to make degenerate PCR, got about 1500 and 150 bp PCR product respectively, they were transformed into the E.coli DH5α through being linked with pMD18-T vector and sequenced after filtration. Similarity alignment showed that the 1500bp products were very similar to the large subunit of hydrogenase 3 genes, and shared 99% identity to the large subunit of hydrogenase 3 from Enterobacter cancerogenus ATCC 35316, and only had the difference of three amino acids; the 150bp products were very similar to the hycA genes, and shared 92% identity to the hycA from Enterobacter cloacae subsp. cloacae ATCC 13047, and only had the difference of four amino acids. Cloning these two fragments would not only enrich the gene resources of hycA and large subunit of hydrogenase 3 genes, but also give the scientific warrant for the metabolic engineering research.