Cloning and preliminary functional analysis of ATPase alpha subunit gene promoter from Dunaliella salina
杜氏盐藻叶绿体ATP 合酶α亚单位基因启动子的克隆及初步功能分析

To clone and identify the function of the promoter of the ATPase alpha subunit (atpA) gene from Dunaliella salina, a chloroplast GenomeWalker DNA Library from D. salina was constructed, from which the 1 347 bp upstream fragment of atpA gene was cloned by long distance-polymerase chain reaction (LD-PCR). According to the positions of the different promoter elements predicted by bioinformatics tools, 5'-deleted series primers were designed to amplify different promoter fragments. Series 5'-deleted expression vector containing the GFP gene was constructed and then transformed to cells of D. salina by particle bombardment. The transformed cells with p1000 recombinant vector produced green fluorescence, but not in other transformation groups, suggesting that the cloned region had the activity of the promoter and may drive the effective expression of the GFP gene in D. salina. Our results have layed a foundation for the chloroplast transformation system of D. salina.