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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 :大肠杆菌表达HPV16E6蛋白的获得可为HPV相关性肿瘤的血清学诊断和流行病学研究打下基础
OBJECTIVE: To investigate the E6 serology of human papillomavirus type 16 associated tumors. Methods: The HPV16E6 gene fragment was amplified from cervical cancer tissue by polymerase chain reaction (PCR), cloned into the sequencing plasmid pGEM T, and the HPV16E6 gene was excised by restriction enzyme and cloned into the expression plasmid pGEMEX 1 , Induced by IPTG as a fusion protein, molecular weight of about 45KD. Results: The fusion protein accounted for 25% of the total bacterial proteins. Western blot analysis showed that HPV16E6 fusion protein could be recognized by anti-HPV16E6 antibody. Conclusion: The expression of HPV16E6 protein in Escherichia coli can provide the basis for serological diagnosis and epidemiological study of HPV-associated tumors