首页 | 本学科首页   官方微博 | 高级检索  
检索        

结核分枝杆菌Ag85B-Ag85A双抗原融合真核表达质粒的构建及表达
引用本文:江山,朱道银,蒋英,骆旭东,陈全.结核分枝杆菌Ag85B-Ag85A双抗原融合真核表达质粒的构建及表达[J].第四军医大学学报,2003,24(21):1973-1975.
作者姓名:江山  朱道银  蒋英  骆旭东  陈全
作者单位:重庆医科大学微生物学教研室,重庆,400016
基金项目:重庆市卫生局重点项目 (0 0 1 0 0 6)
摘    要:目的:构建结核分枝杆菌分泌蛋白Ag85B-Ag85A融合基因及其双价抗原真核表达载体.方法:采用gene SOE-ing法将Ag85B和Ag85A编码基因用疏水甘氨酸接头(Gly4Ser)3行PCR扩增融合,经限制性内切酶消化后克隆入pcDNA3.1( )中构建真核表达质粒pCDAg85B-A,酶切、DNA测序鉴定,用脂质体法将pCDAg85B-A转染COS-7细胞,采用RT-PCR,ELISA方法检测其表达.结果:Ag85B-Ag85A融合基因定向克隆人pcDNA3.1( ),双向测序表明碱基无突变,Ag85B-Ag85A融合基因在真核细胞中能表达.结论:成功构建结核分枝杆菌分泌蛋白Ag85B-Ag85A融合基因及其双价抗原真核表达载体,为结核病基因疫苗的研究奠定基础.

关 键 词:结核分枝杆菌  Ag85B  Ag85A  融合基因  双抗原融合真核表达质粒  基因表达
文章编号:1000-2790(2003)21-1973-03
修稿时间:2003年1月24日

Construction of the fused eukaryotic expression vector of Mycobacterium tuberculosis Ag85B and Ag85A antigens and its expression
JIANG Shan,ZHU Dao Yin,JIANG Ying,LUO Xu Dong,CHEN Quan.Construction of the fused eukaryotic expression vector of Mycobacterium tuberculosis Ag85B and Ag85A antigens and its expression[J].Journal of the Fourth Military Medical University,2003,24(21):1973-1975.
Authors:JIANG Shan  ZHU Dao Yin  JIANG Ying  LUO Xu Dong  CHEN Quan
Institution:JIANG Shan,ZHU Dao Yin,JIANG Ying,LUO Xu Dong,CHEN Quan Department of Microbiology,Chongqing University of Medical Sciences,Chongqing 4000016,China
Abstract:AIM: To construct eukaryotic expression vector of fused gene encoding Mycobacterium tuberculosis Ag85B and Ag85A. METHODS: Fused gene encoding Ag85B and Ag85A of tubercle bacilli was linked with the(Gly 4Ser) 3 linker by gene SOEing. The fused gene was inserted into pcDNA3.1(+). The recombinant plasmid pCDAg85B A was transfected into COS 7 cells with liposome. The expression was detected with RT PCR and ELISA. RESULTS : The fused gene encoding Ag85 and Ag85A was cloned into pcDNA3.1(+) correctly, and no mutation was observed, and its expression was determined in COS 7 cells. CONCLUSION: Eukaryotic recombinant plasmids encoding fused gene of Ag85B and Ag85A antigens were constructed successfully. The results established the basis for further study of the DNA vaccine against tuberculosis.
Keywords:Mycobacterium tuberculosis  Ag85B  Ag85A  fused gene
本文献已被 CNKI 维普 万方数据 等数据库收录!
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