人乳头瘤病毒16型晚期基因L1在大肠杆菌中的表达

ESTABLISHMENT AND IDENTIFICATION OF EXPRESSION SYSTEM OF LATE GENE L1 OF HUMAN PAPILLOMAVIRUS TYPE 16 IN E.coli

人乳头瘤病毒（ＨＰＶ）感染与宫颈癌的发生关系密切。其晚期蛋白Ｌ１是主要结构蛋白，可以刺激机体产生中和抗体，对病毒攻击可以起到保护作用。本研究将ＨＰＶ１６型中国分离株的Ｌ１基因定向克隆到原核表达载体ｐＥＴ－２１ｃ质粒中，建立ＨＰＶ１６ Ｌ１在大肠杆菌中的高效表达系统ｐＥＴ２１ｃ－ＨＰＶ１６Ｌ１，该系统在ＩＰＴＧ的诱导下可表达 ６０ｋｕ的 Ｌ１蛋白，表达效率为 ２３％，此蛋白通过Ｗｅｓｔｅｒｎ－ｂｌｏｔ得到证实。该表达体系的成功建立可以为进一步研究ＨＰＶ１６ Ｌ１蛋白的分子和免疫学特性以及基因疫苗的研制提供实验依据。

Human papillomavirus type 16 (HPV16) is thought to be a very important etiological factor of cervical carcino- ma. The late protein L1 of HPV16 is the main structUral protein, which induce antibody that can neutralize the infection of HPV16. The L1 gene of HPV16 Chinese strain was clamal into plasmid pET-21c. An inducible expression system for HPV16L1 gene in E. coli, pET-21c-HPV16L1 was constructed. Polyacryldride gel electrophoresis showed that pET-21c- HPV16L1 could express L1 protein in size of about 60 KD in E. coli BL21-DE3 under induction of IPTG. The expression effi- ciency was 23 %. The protein was confirmal by western-blot.