摘要
目的 :获得含人乳头瘤病毒 (HPV) 5 8型E6基因的克隆及表达重组体并体外表达E6蛋白 .方法 :聚合酶链方法从 1例宫颈腺癌患者癌组织DNA中获得HPV5 8E6基因 ,并将其与克隆载体pGEM TEasy连接 ,获得重组体HPV5 8 E6 pGEM T ,继之以双酶切将E6基因与同样双酶切的线性化的pRSET A表达载体连接 ,得到E6表达重组体pRSET 5 8E6 ,转化E .coliBL2 1 (DE3) ,用IPTG诱导表达 .结果 :从 1例宫颈癌患者中成功获得了少见HPV5 8型的E6基因并构建了其重组表达载体 .经IPTG诱导后可表达Mr2 4 0 0 0的 6HisHPV5 8E6融合蛋白 ,表达量占菌体蛋白的 1 0 % .结论 :成功获得了少见HPV5 8高危型的E6基因 ,并可在E .
AIM: To obtain human papillomavirus type (HPV) 58 E6 gene and express it in vitro . METHODS: HPV58 E6 gene was amplified from DNA of a cervical cancer patient by PCR, and inserted into pGEM T Easy vector. HPV 58 E6 gene was digested from the recombinant plasmid HPV 58 E6 pGEM T, and ligated into lined prokaryotic expression vector pRSET A with Sac I/ Hin dⅢ, and then expression of 58E6 protein was induced by IPTG. RESULTS: Full length HPV58 E6 gene was successfully amplified from DNA of a cervical cancer patient by PCR. SDS PAGE analysis showed that His6 HPV58 E6 fusion protein of M r 24 000 was expressed after induction of IPTG. The quantity of fusion protein was 10% of total bacterial proteins. CONCLUSION: HPV58 E6 gene is successfully cloned and effectively expressed in E. coli .
出处
《第四军医大学学报》
CAS
北大核心
2004年第3期201-203,共3页
Journal of the Fourth Military Medical University
