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J亚群禽白血病JL-2株gp85基因的克隆与表达
引用本文:宋素泉,付朝阳,高宏雷,王笑梅,曾祥伟,郭艳,王英,张厚双,王晓艳,魏萍.J亚群禽白血病JL-2株gp85基因的克隆与表达[J].中国预防兽医学报,2005,27(5):356-359.
作者姓名:宋素泉  付朝阳  高宏雷  王笑梅  曾祥伟  郭艳  王英  张厚双  王晓艳  魏萍
作者单位:1. 东北农业大学
2. 中国农业科学院哈尔滨兽医研究所,黑龙江,哈尔滨,150001
摘    要:本实验在分离鉴定J亚群禽白血病JL-2株的基础上,利用PCR方法扩增出包括gp85基因在内的长度为960bp的DNA片段.将其连到PGEX-6p-1载体上,构建了重组表达质粒PGEX-6P-1/gp85,对质粒进行序列分析并在IPTG的诱导下进行表达.SDS-PAGE分析结果表明,融合蛋白(54 Ku)在BL21中得到有效表达.表达产物占菌体总蛋白的31.8%.Western Blot结果表明,表达产物可与ALV-J阳性血清发生特异性反应.这些结果为进一步研究gp85蛋白的功能及研制ALV-J ELISA诊断试剂盒奠定了基础.

关 键 词:J-亚群禽白血病病毒  gp85基因  克隆  表达
文章编号:1008-0589(2005)05-0356-04
收稿时间:2004-04-05
修稿时间:2004年4月5日

Cloning and expression of gp85 gene of avian leukosis virus subgroup J JL-2 strain
SONG Su-quan,FU Chao-yang,GAO Hong-lei,WANG Xiao-mei,ZENG Xiang-wei,GUO Yan,WANG Ying,ZHANG Hou-shuang,WANG Xiao-yan,WEI Ping.Cloning and expression of gp85 gene of avian leukosis virus subgroup J JL-2 strain[J].Chinese Journal of Preventive Veterinary Medicine,2005,27(5):356-359.
Authors:SONG Su-quan  FU Chao-yang  GAO Hong-lei  WANG Xiao-mei  ZENG Xiang-wei  GUO Yan  WANG Ying  ZHANG Hou-shuang  WANG Xiao-yan  WEI Ping
Abstract:A DNA fragment encoding gp85 of Avian Leukosis virus subgroup J JL-2 strain was amplified by PCR and cloned into PGEX-6P-1 expression plasmid.After sequencing,the recombined plasmid was transformed into E.coli BL21(DE3) and induced by IPTG.SDS-PAGE analysis showed that the fusion protein was efficiently expressed.The result of Western Blotting also showed that the fusion protein expressed could react specifically with the ALV-1 positive serum.
Keywords:avian leukosis virus subgroup J  gp85 gene  cloning  expression
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