宫颈癌组织中HPV16E6的基因克隆及其在大肠杆菌中的表达

GENE CLONING AND EXPRESSING IN PROKARYOTIC CELL OF HPV16 E6 FROM THE CERVICAL CANCER

目的 :进行人乳头瘤病毒 16型相关肿瘤的E6血清学研究。方法 :采用聚合酶链反应 (PCR)从宫颈癌组织DNA中扩增出HPV16E6基因片段 ,克隆至测序质粒 pGEM T中 ,并用限制性内切酶将HPV16E6基因切下 ,克隆至表达质粒pGEMEX 1中 ,经IPTG诱导表达为融合蛋白 ,分子量约 4 5KD。结果 :融合蛋白占菌体总蛋白的 2 5% ,免疫印迹检测表明HPV16E6融合蛋白能被抗HPV16E6抗体识别。结论

Objective:To deepen serological study of E6 in tumors associated with human papillomavirus 16 type (HPV16).Methods:PCR technique was used to amplify the HPV16E6 gene fragment from the DNA of cervical cancer tissue. The fragment was then cloned into the pGEMT.After being digested with SalI and NotI, the HPV16E6 was cloned into the expression vector pGEMEX1. The recombinant vector was induced by IPTG to express the fusion protein with molecular weight of 45KD.Results:The fusion protein accounted for 25%of the whole protein and the expression protein was recognized by the antiE6 antibody in the Western Blot.Conclusion:This protein can be used as a basic antigen in the serological diagnosis of HPVassociated tumors.