Cloning and Characterization of atpA Gene Fragment from Chloroplast of Dunaliella salina

摘要

According to conserved motif of the homologous amino acid sequence of about ten kinds of algae, we designed a pair of degenerate primer and amplified atpA gene fragment from the chloroplast of Dunaliella salina by PCR technique.The resulting PCR products were inserted into pMD-18 T-vector,and then transformed into E.coli JM109. Positive colonies were selected to determine their sequence. Homologous analysis of the deduced amino acid sequence were performed by BLAST and subsequently compared with GenBank data.Use the fragment we cloned as prob and hybridize with chloroplast DNA of D.salina by southern blot method. The nucleotide sequences were 405bp which encode 135 amino acid. The sequence shared high homology with atpA, with identity 92%, 88%, 87%, 86% and 85% to Chlamydomonas reinhardtii, Chlorella vulgaris, Mesostigma viride, Nephroselmis olivacea and Cyanidioschyzon merolae respectively.Southern blot result showed that there were obervious hybridization signals on cpDNA of D.salina. It can be concluded that the cloned sequence is atpA gene fragment from the chloroplast of Dunaliella salina.