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目的:构建人乳头瘤病毒16L1的原核表达质粒,为下一步探讨蛋白的表达以及蛋白的功能研究作准备。方法:以临床HPV16阳性标本为模板,PCR扩增L1的片段,L1片段经EcoRⅠ和SalⅠ双酶切后,插入载体pGEX4T-1,转化JM109感受态细胞,平板筛选获得pGEX4T-1/HPV L1的阳性质粒。通过酶切、测序验证质粒的正确性。结果:构建了原核表达质粒pGEX4T-1/HPV16L1,并通过酶切、测序等方法验证其完全正确。结论:成功构建的pGEX4T-1/HPVL1为今后的蛋白的表达和功能研究打好了坚实的基础,为HPV16的疫苗研究提供了有益的支持。
OBJECTIVE: To construct a prokaryotic expression plasmid of human papillomavirus 16L1 in preparation for further study of protein expression and protein function. Methods: The HPV16 positive samples were used as template to amplify the L1 fragment. The L1 fragment was double digested with EcoRⅠ and SalⅠ and inserted into the vector pGEX4T-1 to transform into competent cells of JM109. The pGEX4T-1 / HPV L1 Positive plasmids. The plasmid was verified by restriction enzyme digestion and sequencing. Results: The prokaryotic expression plasmid pGEX4T-1 / HPV16L1 was constructed and verified by restriction enzyme digestion and sequencing. Conclusion: The successful construction of pGEX4T-1 / HPVL1 lay a solid foundation for the future study of protein expression and function and provide useful support for the vaccine research of HPV16.