| 人乳头瘤病毒16亚型L1蛋白在多形汉逊酵母中的优化表达 |
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| 引用本文： | 李巍巍,何秀萍,郭雪娜,张振颖,张博润. 人乳头瘤病毒16亚型L1蛋白在多形汉逊酵母中的优化表达[J]. 生物工程学报, 2009, 25(10): 1516-1523 |
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| 作者姓名： | 李巍巍 何秀萍 郭雪娜 张振颖 张博润 |
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| 作者单位： | 1. 中国科学院微生物研究所酵母菌分子遗传与育种实验室,北京,100101;中国科学院研究生院,北京,100049
2. 中国科学院微生物研究所酵母菌分子遗传与育种实验室,北京,100101 |
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| 摘 要： | 为了实现人乳头瘤病毒(Humanpa pillomavirus,HPV)16亚型衣壳蛋白L1在多形汉逊酵母(Hansenula polymorpha)中的高效表达,根据L1蛋白的氨基酸序列及多形汉逊酵母的密码子偏爱性,对L1蛋白的编码序列进行优化设计,合成了完整的编码序列,命名为HPV16L1。以甲醇诱导型启动子MOXp和终止子AOXTT为表达调控元件,以尿嘧啶合成相关基因URA3为筛选标记,构建了HPV16L1的重组表达质粒pYMOXU-HPV16。用SacII酶切质粒pYMOXU-HPV16使其线性化,电转化多形汉逊酵母菌株H-ura3,依据营养缺陷互补筛选重组菌株。通过PCR扩增及HPV16L1蛋白表达量分析表明已获得稳定高表达L1蛋白的重组汉逊酵母菌株HP-U-16L。摇瓶发酵条件的初步优化表明,以YPM(pH7.0)为基础培养基进行诱导培养,控制接种量使初始培养液OD600为1.0,每隔12h补加甲醇至终浓度为1%(V/V),37oC、200r/min条件下诱导培养72h后,HPV16L1蛋白的最高表达量为78.6mg/L。本研究为多形汉逊酵母源HPV16L1疫苗的研制奠定了基础。
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| 关 键 词： | 人乳头瘤病毒16亚型 衣壳蛋白L1 多形汉逊酵母 优化表达 |
| 收稿时间： | 2009-07-06 |
Optimized expression of the L1 protein of human papillomavirus in Hansenula polymorpha |
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| Weiwei Li,Xiuping He,Xuena Guo,Zhenying Zhang and Borun Zhang. Optimized expression of the L1 protein of human papillomavirus in Hansenula polymorpha[J]. Chinese journal of biotechnology, 2009, 25(10): 1516-1523 |
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| Authors: | Weiwei Li Xiuping He Xuena Guo Zhenying Zhang Borun Zhang |
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| Affiliation: | Laboratory of Yeast Molecular Genetics and Breeding, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; Graduate University of Chinese Academy of Sciences, Beijing 100049, China;Laboratory of Yeast Molecular Genetics and Breeding, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;Laboratory of Yeast Molecular Genetics and Breeding, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;Laboratory of Yeast Molecular Genetics and Breeding, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;Laboratory of Yeast Molecular Genetics and Breeding, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China |
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| Abstract: | The heterologously expressed L1 protein of human papilomavirus 16 can assembly into virus-like particles (VLPs), which has been used as prophylactic vaccine for cervical carcinoma. To express L1 protein in Hansenula polymorpha, we analyzed the codon usage of the native gene of L1 protein and redesigned the encoding sequence according to the codon bias of H. polymorpha. We used assembly PCR to synthesize the native gene HPV16L1-N and the codon optimized gene HPV16L1. The synthesized genes were cloned into pM... |
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| Keywords: | human papilomavirus 16 L1 protein Hansenula polymorpha optimized expression |
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