人乳头瘤病毒16型E7基因的克隆和表达

SUBCLONING AND EXPRESSION OF HUMAN PAPILLOMAVIRUS TYPE 16 E7 GENE IN ESCHERICHIA COLI

用分子克隆技术在大肠杆菌中表达HPV16 E7基因。用限制性内切酶DdeⅡ将含有大部分E7基因的191bp片段从HPV16基因组中切出。将此片段与含有色氨酸操纵子的启动子的表达载体pATH10连接成重组DNA,用以转化E.coliDH5a,并筛选出阳性重组子。还进行了限制酶酶切分析和蛋白质SDS-PAGE分析。表达的新蛋白质带的分子量与预期的分子量相符。我们认为表达的新蛋白质是TrpE和E7蛋白的融合蛋白。

The E7 gene of human papillomavirus type 16 was expressed in E. coli with molecular cloning technique. A I91 bp fragment containing most of the E7 gene was cut out from the HPV 16 genome with the restriction enzyme Dde Ⅰ. The plasmid pATH10 which contains the promoter of the tryptophane operon was chosen as the expression vector. The 191 bp fragment and the vector plasmid were ligated together later and the recombinant DNA formed was used to transform E. coli DH5α. Restriction enzyme mapping and SDS-PAGE protein analysis were done on four transformants. The molecular weight of the newly expressed protein of the transformant was estimated and found to correspond well with that expected theoretically. We concluded that the expressed protein was a fusion protein of TrpE/E7_(32-95aa).