[HPV31和52 L2融合蛋白在大肠杆菌中的高表达及其免疫原性检测]。

中华实验和临床病毒学杂志 Pub Date : 2013-04-01
Ling Zhou, Jiao Ren, Li Zhao, Jing Feng, Ming-Qiang Hao, Wen-Jie Tan, Li Ruan, Peng-Peng Qu, Hou-Wen Tian
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引用次数: 0

摘要

目的:表达HPV31与52 L2融合蛋白并检测其免疫原性。方法:根据GenBank数据库中公布的HPV31和52 L2 11-200AA的氨基酸序列,人工合成根据大肠杆菌密码子使用情况优化的HPV31和52 L2融合基因,编码HPV31和HPV52 L2的11-200个氨基酸,并克隆到pET-9a载体上。在原核表达系统中表达了HPV31与52 L2融合蛋白,纯化后用该融合蛋白免疫小鼠。免疫原性在接种小鼠中得到表征。结果:HPV31和52 L2融合蛋白在大肠杆菌中高表达,融合蛋白的表达量接近细菌总蛋白的20%。纯化的铝佐剂融合蛋白可诱导ELISA检测特异性高滴度的IgG抗体,并可诱导HPV31、HPV52假病毒的中和抗体和HPV45、58、16、18假病毒的交叉中和抗体。结论:HPV31和52 L2融合蛋白可诱导抗HPV假病毒的中和抗体和交叉中和抗体。为研制HPV L2蛋白疫苗提供了实验室依据。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[The high expression of HPV31 and 52 L2 fusion protein in E. coli and detection of its immunogenicity].

Objective: To express HPV31 and 52 L2 fusion protein and detect its immunogenicity.

Methods: According to the amino acid sequences of HPV31 and 52 L2 11-200AA published in the GenBank database, weartificially synthesized the HPV31 and 52 L2 fusion gene which was optimized according to Escherichia coli codon usage and encodes 11-200 amino acid of HPV31 and HPV52 L2, then cloned it into pET-9a vector. The HPV31 and 52 L2 fusion protein was expressed in Prokaryotic expression system and the mice were immunized with the fusion protein after purification. The immunogenicity was characterized in vaccinated mice.

Results: HPV31 and 52 L2 fusion protein was highly expressed in E. coli, the amount of fusion protein is nearly 20% of the total bacterial protein. The purified fusion protein with aluminum adjuvant could induce specific high titer of IgG antibodies detected by ELISA, and also induce the neutralizing antibodies against pseudovirus of HPV31 and HPV52 and cross-neutralizing antibodies against pseudovirus of HPV45, 58, 16, 18.

Conclusion: HPV31 and 52 L2 fusion protein could induce neutralizing and cross-neutralizing antibodies against HPV pseudovirus. It provides laboratory basis for development of HPV L2 protein vaccine.

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