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高危人乳头瘤病毒58型E6基因的克隆及表达

张菊,陈中灿,白玉杰,高艳娥,何玉宪,阎小君

Journal of the Fourth Military Medical University（2004）

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Abstract
目的:获得含人乳头瘤病毒（HPV）58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HPV58 E6基因，并将其与克隆载体pGEM-T Easy连接，获得重组体HPV58-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-58E6，转化E.coli BL21(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HPV58型的E6基因并构建了其重组表达载体. 经IPTG诱导后可表达Mr 24 000的6His HPV58 E6融合蛋白，表达量占菌体蛋白的10%. 结论：成功获得了少见HPV58高危型的E6基因，并可在E.coli中高效表达.
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Key words
human papillomavirus type,e6 gene,cloning
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