鱼腥藻PCC7120基因asr0757/alr0758的初步研究

A Primary Study of Genes asr0757/alr0758 in Anabaena sp. PCC7120

为研究鱼腥藻PCC7120(Anabaena sp.)基因asr0757/alr0758在毒素-抗毒素系统中的相关生物学功能,设计了特异性引物,扩增目的片段asr0757和alr0758,将目的基因与p MD18-T载体连接构建克隆载体,并对其进行XhoⅠ和NdeⅠ双酶切,再与表达载体p ET-28a连接构建表达重组菌,重组菌的体外表达在IPTG的诱导下进行。经琼脂糖电泳检测,结果扩增出了大小为210 bp的asr0757和342 bp的alr0758目的基因。经SDS-PAGE电泳检测,表达出相对分子质量分别为8.068 k D和12.534 k D的蛋白质。根据结果可初步认定alr0758为毒素基因,asr0757为抗毒素基因,共同构成鱼腥藻PCC7120毒素-抗毒素系统。

For the study on the related biological function of Anabaena sp. PCC7120 gene asr0757 / alr0758 in toxin-antitoxin system, the specific primers were designed. The target fragments of genes asr0757 / alr0758 were amplified by PCR. Their products were then inserted into p MD18-T vector after the XhoⅠ and NdeⅠ double enzyme digestion and recombinant fungus was structured by connecting the p ET-28 a vector, with its expression in vitro guided by IPTG. The results showed that objective gene asr0757 was 210 bp and alr0758 was 342 bp by agarose gel electrophoresis detection. Their protein molecular weight were 8.068 k D and 12.534 k D by SDS-PAGE electrophoresis detection. It can be preliminarily determined that alr0758 was the toxin gene,while asr0757 was the antitoxin gene. Both of them constituted a toxin-antitoxin system.