人乳头瘤病毒6型L1基因的克隆及蛋白表达

目的：在大肠杆菌中表达经密码子优化的人乳头瘤病毒6型（HPV6）L1的融合蛋白。方法：PCR方法扩增HPV6 L1，基因，测序及序列比对后，对基因进行密码子优化并合成优化后的基因HPV6mLI，将其克隆入原核表达载体pGEX4T-1，IPTG诱导融合蛋白在大肠杆菌BL21（DE3）中表达，SDS-PAGE鉴定表达产物。结果：酶切和测序结果证实HPV6 mL1基因的原核表达载体构建正确；以1mmol／L IPTG于37℃诱导4h，蛋白以包涵体形式表达；表达产物的相对分子质量与预期值一致，为80000。结论：获得大肠杆菌表达的HPV6L1蛋白，为其结构功能研究和疫苗研发提供了基础。

Cloning and Expression of Human Papillomavirus Type 6 L1 Gene

Objective： To express optimized-code fusion protein L1 of human papillomavirus type 6 （HPV6） in E.coli BL21 （DE3）. Methods： HPV6 L1 gene was amplified by PCR and the optimized-code HPV6 mL1 gene was synthesized basing on sequencing and sequence comparison. The HPV6 roLl gene was inserted into the prokaryotic vector pGEX4T-1 to construct the expression plasmid, and then the HPV6 taLl protein was expressed after induction of IPTG in E.coli BL21（DE3）. The fusion protein was determined by SDS-PAGE. Results： Restriction enzyme digestion analysis and DNA sequencing showed that the prokaryotic expression system of HPV6 mL1 gene was constructed successfully. When induced by 1 mmol/L IPTG lasted 4 h at 37℃, fusion protein HPV6 L1 was expressed in inclusion body form. SDS-PAGE showed that the molecular weight of fusion protein was 80 kD and was identical with predict size. Conclusion： Fusion protein HPV6 L1 was obtained, which provides a basis for further vaccine and functional assay.