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人乳头瘤病毒6型L1基因的克隆及蛋白表达
引用本文:刘斌,钱永华,张骞,张万菊,张钫,麻粉莲,郑丽舒. 人乳头瘤病毒6型L1基因的克隆及蛋白表达[J]. 生物技术通讯, 2009, 20(4): 495-497. DOI: 10.3969/j.issn.1009-0002.2009.04.012
作者姓名:刘斌  钱永华  张骞  张万菊  张钫  麻粉莲  郑丽舒
作者单位:1. 西北农林科技大学,动物医学院,陕西,杨凌,712100
2. 中国疾病预防控制中心,病毒病预防控制所,北京,100052
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。

关 键 词:人乳头瘤病毒6型  L1基因  蛋白表达

Cloning and Expression of Human Papillomavirus Type 6 L1 Gene
LIU Bin,QIAN Yong-Hua,ZHANG Qian,ZHANG Wan-Ju,ZHANG Fang,MA Fen-Lian,ZHENG Li-Shu. Cloning and Expression of Human Papillomavirus Type 6 L1 Gene[J]. Letters in Biotechnology, 2009, 20(4): 495-497. DOI: 10.3969/j.issn.1009-0002.2009.04.012
Authors:LIU Bin  QIAN Yong-Hua  ZHANG Qian  ZHANG Wan-Ju  ZHANG Fang  MA Fen-Lian  ZHENG Li-Shu
Affiliation:1. College of Veterinay Medicine, Northwest A&F University, Yangling 712100;2. National Institute for Viral Disease Control and Prevention, China CDC, Beijing 100052; China)
Abstract: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.
Keywords:human papillomavirus type 6  L1 gene  protein expression
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