Cloning and structure analysis of hydrogenase gene from Chlamydomonas reinhardtii SE

The cDNA of Chlamydomonas reinhardtii SE encoding hydrogenase (HydA2) was obtained from the total RNA of C. reinhardtii SE by RT-PCR. The DNA of hydrogenase was amplified by PCR from the genomic DNA of C. reinhardtii SE. The cDNA and DNA of hydrogenase were sequenced, respectively. The structure of hydrogenase gene was analyzed by biology software. The open reading frame predicts that the hydrogenase is composed of 3584 bp encoding 505 amino acids in length with a predicted M.W. of 53.69 kDa. Ten exons (including 1518 bp) and nine introns (including 2066 bp) have been found in the hydrogenase, and there were two potential N-glycosylate sites, eight protein kinase C phosphorylation site, eight casein kinase II phosphorylation site and one sulphorylation in the sequence. The theory pI was 6.15. Total number of negatively charged residues (Asp + Glu) and positively charged residues (Arg + Lys) were 55 and 61, respectively.