【摘要】 目的:利用基因重组技术,构建人乳头瘤病毒18型(HPV18)L1基因果蝇表达质粒。方法:以pUC18-HPV18为模板,运用PCR方法扩增不含核定位序列的HPV18L1DNA片段;PCR产物连接至PGEM-TEasy质粒,并测序鉴定;将测序正确的HPV18L1DNA片段亚克隆到果蝇表达载体pMT/BiP/V5-HisA中;利用酶切及PCR方法鉴定重组质粒。结果:经双酶切及PCR鉴定,证实成功构建重组果蝇表达质粒pMT/BiP/V5-HPV18L1。结论:pMT/BiP/V5-HPV18L1重组果蝇表达质粒为下一步转染果蝇S2细胞及制备HPV18VLPs奠定了基础。更多还原

【Abstract】 Objective: To construct recombinant HPV18L1 expression system in Drosophila S2 cells. Methods: The truncated HPV18 L1 gene fragment (no nuclear-localization sequence)was amplified by PCR from pUC18-HPV18 plasmid ;PCR products were inserted into the pGEMT-Easy vector and identified by sequencing, then the truncated HPV18L1 fragment was cloned into Drosophila Schneider expression vector pMT/BiP/V5-HisA, to construct pMT/BiP/V5-HisA/ HPV18L1 plasmid;The construct was identified with endonuclease cleavage and PCR amplification . Results: the truncated HPV18L1 fragment was correctively amplified and successfully cloned in to pMT/BiP/V5-HisA expression plasmid. Conclusion: The construct of pMT/BiP/V5-HPV18L1 may provide a prerequisite for expression of HPV18L1 VLPs in Drosophila S2 cells in the future.更多还原