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猪瘟病毒E2基因在Pichia pastoris中的表达及其免疫活性的初步研究
引用本文:韩雪清,刘湘涛,张涌,谢庆阁,田波.猪瘟病毒E2基因在Pichia pastoris中的表达及其免疫活性的初步研究[J].生物工程学报,2002,18(2):208-211.
作者姓名:韩雪清  刘湘涛  张涌  谢庆阁  田波
作者单位:1. 中国农业科学院兰州兽医研究所农业部畜禽病毒病重点实验室,兰州,730046
2. 西北农林科技大学,杨凌,712100
3. 中国科学院微生物研究所,北京,100080
基金项目:国家重大基础研究发展规划项目 (No .G19990 119)~~
摘    要:将猪瘟病毒的E2基因克隆入酵母分泌型表达载体pPIC9K中,酶切线性化后电穿孔导入Pichia pastoris进行整合,经G418筛选得到高拷贝转化子,甲醇诱导表达。SDS-PAGE和Western blit结果证实了酵母培养上清液中含有E2蛋白。免疫活性研究证明P.pastoris表达的E2蛋白能刺激动物产生抗猪瘟病毒的抗体。

关 键 词:猪瘟病毒  E2基因  毕赤酵母  表达  免疫活性
文章编号:1000-3061(2002)02-0208-04
修稿时间:2001年9月10日

Study on the Expression of E2 Gene of Classical Swine Fever Virus in Pichia pastoris and the Immunological Activity of Its Expression Product
HAN Xue Qing,LIU Xiang Tao,ZHANG Yong,XIE Qing Ge,Tian Bo.Study on the Expression of E2 Gene of Classical Swine Fever Virus in Pichia pastoris and the Immunological Activity of Its Expression Product[J].Chinese Journal of Biotechnology,2002,18(2):208-211.
Authors:HAN Xue Qing  LIU Xiang Tao  ZHANG Yong  XIE Qing Ge  Tian Bo
Institution:Lanzhou Veterinary Research Institute, CAS, Lanzhou 730046, China. hanxq8@hotmail.com
Abstract:E2 gene of classical swine fever virus (CSFV) was cloned into secretory pPIC9K Pichia pastoris expression vector. After being linearized by digestion , the vector was transformed into Pichia pastoris by electroporation to integrate with the genome, the transformants with high copies were screened by G418 and were induced to express with methonal. The results of SDS PAGE and Western blot demonstrated that the supernatant of the induced P.pastoris culture contained protein E2. The results of the study on the immunological activity indicated that the protein E2 expressed in P.pastoris can elicit animal bodies to produce antibodies against protein E2.
Keywords:classical swine fever virus  E2 gene    Pichia pastoris    expression  immunological activity
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