摘要
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HPV6L1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HPV6 mL1,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。
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.
出处
《生物技术通讯》
CAS
2009年第4期495-497,共3页
Letters in Biotechnology
