鱼腥藻PCC7120α质粒上的parD/E同源基因all7155/asl7156的克隆及表达

Cloning and Expressing of Homologous Gene all7155 / asl7156 of parD/E in α Plasmid of Anabaena sp. PCC 7120

为研究鱼腥藻PCC7120质粒上毒素抗毒素系统的蛋白质性质和相互作用,根据NCBI中PCC7120的α质粒上的all7155和asl7156基因数据,通过降落PCR克隆了目的基因(all7155/336 bp,asl7156/258 bp).将目的基因片段连接至p MD18-T构建了克隆载体,蓝白斑筛选了阳性克隆,经双酶切纯化后将目的基因连接至表达载体p ET30a(+),并转入表达菌BL21中,测序后证实构建成功,并利用IPTG诱导进行了表达.通过NCBI的BLAST蛋白比对,发现了all7155和asl7156与已知的Par D/E毒素抗毒素系统同源,可通过对此基因的克隆,表达和蛋白性质来进一步认识Par D/E系统.

In order to research the characteristics and interactions of the toxin-antitoxin protein in the αplasmid of Anabaena sp.PCC7120 , the target genes（all7155/336 bp,asl7156/258 bp） were successfully cloned by touchdown PCR based on the data of all7155/asl7156 inαplasmid of Anabaena sp.PCC 7120 in NCBI.The target genes were connected with the pMD-18T vector to recombine with the clone vector .After the blue-white screening and double enzyme cutting, the expressing pMD-30a（＋） vector was recombined with the target genes and transduced into the expression bacteria BL21. The construction was then confirmed successfully by sequencing and expressed by IPTG induction.According to the protein BLAST of NCBI, the all7155/asl7156 were founded homologous with the known parD/E toxin-antitoxin system .The research on the clone, expression and protein characteristics of the genes could be useful for the understanding of the parD/E system.