首页 | 本学科首页   官方微博 | 高级检索  
   检索      

截短型人乳头瘤病毒58型L1蛋白的表达及其体外生物活性研究
引用本文:李文生,郑瑾,刘红莉,陈宏伟,杨军,王一理,司履生.截短型人乳头瘤病毒58型L1蛋白的表达及其体外生物活性研究[J].生物工程学报,2004,20(4):536-539.
作者姓名:李文生  郑瑾  刘红莉  陈宏伟  杨军  王一理  司履生
作者单位:1. 西安交通大学生命科学与技术学院癌症研究所,环境与疾病相关基因教育部重点实验室,西安,710061;西安交通大学陕西临床学院,西安,710068
2. 西安交通大学生命科学与技术学院癌症研究所,环境与疾病相关基因教育部重点实验室,西安,710061
基金项目:国家自然科学基金资助项目 (No .3 0 2 71184)~~
摘    要:利用PCR技术克隆截短型HPV58 L1基因并重组入杆状病毒表达系统穿梭质粒pFastBac-Htb,通过转座反应,将目的基因片段重组入杆状病毒基因组,分离重组的Bacmid DNA, 并转染Sf-9昆虫细胞,收集被转染的Sf-9细胞,提取细胞蛋白,SDS-PAGE检测可见在大约58Kda处出现一新生蛋白条带,Western blot 证实为HPV58L1蛋白。用ProBondTM纯化系统纯化所表达的蛋白。小鼠红细胞凝集试验证实纯化的蛋白可介导小鼠红细胞凝集,透射电镜观察证实纯化蛋白可自组装成VLP。结果表明昆虫杆状病毒表达系统可高效表达截短型HPV58L1蛋白,纯化后的截短型HPV58L1蛋白在体外可自组装VLP,并具有介导小鼠红细胞凝集的生物学活性。

关 键 词:截短型HPV58  L1蛋白,  昆虫杆状病毒表达系统,  蛋白纯化,  病毒样颗粒
文章编号:1000-3061(2004)04-0536-04
修稿时间:2003年12月29

Carboxyl Terminus Truncated HPV58 Virus L1 Protein Expressed with Baculovirus System and its Bioactivity
Wen-Sheng Li,Jin Zheng,Hong-Li Liu,Hong-Wei Chen,Jun Yang,Yi-Li Wang,Lü-Sheng Si.Carboxyl Terminus Truncated HPV58 Virus L1 Protein Expressed with Baculovirus System and its Bioactivity[J].Chinese Journal of Biotechnology,2004,20(4):536-539.
Authors:Wen-Sheng Li  Jin Zheng  Hong-Li Liu  Hong-Wei Chen  Jun Yang  Yi-Li Wang  Lü-Sheng Si
Institution:Institute for Cancer Research School of Life Science & Technology, Xi'an Jiaotong University, Xi'an 710061, China.
Abstract:To prepare carboxyl terminus truncated human papillomavirus type 58L1 protein, and study on its in vitro bioactivity. PCR was used to amplify carboxyl terminus truncated HPV 58L1 gene, the product was inserted into the PUCMT cloning vector, preparing recombinant PFastBacHTb containing carboxyl terminus truncated HPV58L1 gene. Further more, the recombinant plasmid PfastbacHTb was used to transform DH10Bac cells, constructing recombinant Baculovirus, then the recombinant virus was successfully used to infect Sf-9 insect cells. After incubating at 27 degrees C for 72 hours, the infected cells were collected and total cellular proteins were extracted. The target protein with MW 58KD was revealed by SDS-PAGE and confirmed by Western blot. The interested protein was purified by ProBond purification system. The purified interested protein was identified to self-assemble into VLPs by Transmission electron microscope, and induce murine erythrocyte hemagglutination, indicating that the given proteins had the conformation of VLPs, collecting, HPV58L1 proteins with carboxyl terminus truncation could be efficiently expressed in baculovirus Sf-9 cells expression system, it has identical in vitro bioactivity to the wild type HPV58L1, The present study is fundmental for preparing HPV58L1 prophylactic vaccine.
Keywords:Hpv58 L1protein of carboxyl terminus truncation  baculovirus expression system  protein purification  virus-like particles
本文献已被 CNKI 维普 万方数据 等数据库收录!
点击此处可从《生物工程学报》浏览原始摘要信息
点击此处可从《生物工程学报》下载免费的PDF全文
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号