结核分枝杆菌融合抗原Ag85B-ESAT6真核表达载体的构建及鉴定  

CONSTRUCTION AND IDENTIFICATION OF EUKARYOTIC VECTOR EXPRESSING MYCOBACTERIUM TUBERCULOSIS FUSION ANTIGEN AG85B-ESAT6

作　　者：程春明[1] 曹以诚[1] 杜正平[1] 杨化强[1] 郭旭[1] 方翔[1] 张珍武[1] 

机构地区：[1]华南理工大学生物科学与工程学院,广东广州510640

出　　处：《河南工业大学学报（自然科学版）》2006年第3期29-32,共4页Journal of Henan University of Technology：Natural Science Edition

基　　金：国家自然科学基金重大研究计划项目(90412015);华南理工大学高水平大学建设重大项目(321D75010)

摘　　要：以结核分枝杆菌H37Rv株的基因组作为模板,将Ag85B和ESAT6编码基因进行PCR扩增,然后采用基因剪接重叠扩增PCR法(gene SOEing)将其通过疏水甘氨酸接头(GGIGIAPG)连接融合,定向克隆至质粒Pvax1中,构建结核分枝杆菌Ag85B-ESAT6融合抗原的真核表达质粒Pvax1/AE.经单双限制性内切酶图谱、PCR产物及DNA测序分析等多种方法鉴定,证实Pvax1/AE真核表达质粒构建成功.为进一步研究其结核核酸疫苗原型的免疫保护效果奠定了基础.To construct eukaryotic expression vector expressing Mycobacterium tuberculosis fusion antigen Ag85B- ESAT6, the genes respectively encoding Ag85B and ESAT6 protein were amplified by polymerase chain reaction （PCR） based on genome of MTB H37Rv strain. Then by gene SOEing method, fusion gene encoding Ag85B-ESAT6 protein was linked with the Iinker（GGIGIAPG）. The fusion gene was cloned into pVAX1 . Using single and double restriction endonuclease digestion, PCR and DNA sequencing, it was confirmed that the recombinant eukaryotic expression plasmid （pVAX1/AE） had been successfully constructed. The result serves as a prototype of nucleic acid vaccine for further studies on its immunity effectiveness against MTB.

关 键 词：结核 AG85B ESAT6 融合基因 核酸疫苗 

分 类 号：Q781[生物学—分子生物学]