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林道红, 凌虹, 庄敏, 郭淑元, 马培林, 谷鸿喜. 人乳头瘤病毒16型L1基因重组表达质粒的构建及在E.Coli宿主中的表达与鉴定[J]. 中国公共卫生, 2000, 16(10): 875-876. DOI: 10.11847/zgggws2000-16-10-04
引用本文: 林道红, 凌虹, 庄敏, 郭淑元, 马培林, 谷鸿喜. 人乳头瘤病毒16型L1基因重组表达质粒的构建及在E.Coli宿主中的表达与鉴定[J]. 中国公共卫生, 2000, 16(10): 875-876. DOI: 10.11847/zgggws2000-16-10-04
Lin Daohong, . Expression and Identification of L1 Gene of Human Papilloma Virus Type 16 in E.coli[J]. Chinese Journal of Public Health, 2000, 16(10): 875-876. DOI: 10.11847/zgggws2000-16-10-04
Citation: Lin Daohong, . Expression and Identification of L1 Gene of Human Papilloma Virus Type 16 in E.coli[J]. Chinese Journal of Public Health, 2000, 16(10): 875-876. DOI: 10.11847/zgggws2000-16-10-04

人乳头瘤病毒16型L1基因重组表达质粒的构建及在E.Coli宿主中的表达与鉴定

Expression and Identification of L1 Gene of Human Papilloma Virus Type 16 in E.coli

  • 摘要: 构建pGEX4T1HPV16L1重组表达系统,为进一步研制基因工程疫苗奠定基础,采用PCR方法扩增HPV16中国分离株L1外源基因片段,pGEX4T-1为表达载体,构建pEGX4T-1-HPV16L1重组表达系统,在大肠杆菌宿主BL(21)中,经IPTG诱导,表达融合蛋白GST-L1,经SDSPAGE电泳和Westernblot进行鉴定结果表明,成功构建pEGX4T-1-HPV16L1重组表达质粒并获得高效表达并经Westernblot鉴定为L1外源蛋白.提示:所构建的pGEX4T-1-HPV16L1重组表达质粒能够高效表达HPV16L1蛋白.

     

    Abstract: Ithas been established that the infection of human papilloma virus type 16(HPV16)has highly colse relation with the occurring of women cervical carcinoma.And ithas great practical significance to explore HPV16 genetic vaccine which can prevent the infection of the same type virus.Method PCR method was applied to amplify L1 gene fragment from HPV16L1 chinese isolate.The recombinant p GEX4 T-1-HPV16L1 was constructed from L1 gene fragment and the p GEX4 T-1 vector.After transducting to host BL-1,IPT G inducing,L1 protein was identified by SDS-PA GE and Western blot.Results The recombinant p GEX4 T-1-HPV16L1 was successfully identified and L1 protein was expressed with high quantity.Conclusion p GEX4 T-1-HPV16L1 could express L1 protein with high production which was identified through western blot identification.

     

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