一种结核杆菌融合抗原的克隆表达及活性测定  

The cloning,expression and antigenicity of the fused antigen of Mycobacterium tuberculosis

作　　者：宋晓国[1] 陈坤[1] 冯晓燕[1] 王国华[1] 朱翠侠[1] 李惠文[2] 韦攀健[2] 张贺秋[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850 [2]北京市结核病胸部肿瘤研究所,北京101149

出　　处：《中国实验诊断学》2008年第5期569-573,共5页Chinese Journal of Laboratory Diagnosis

基　　金：国家863重大专项2006AA02090804;国家自然科学基金30772067

摘　　要：目的进一步分析优化结核杆菌抗原优势表位,获得高特异性融合抗原,以满足研制结核病检测试剂的需要。方法用分子生物学软件分析结核杆菌四种抗原MPT64、Mtb8、Mtb8.4和Mtb16.3的优势表位后,分别插入到pBVIL1载体中,构建表达质粒,并利用pBVIL1载体特点,将其基因连接后,进行融合表达,经纯化获得抗原,用间接ELISA法测其活性。结果各抗原优势肽段及融合抗原均获得了高效表达,融合抗原的特异性和灵敏性均高于各单一肽段。结论此融合抗原可用于研制结核病的诊断试剂。Objective In order to obtain the fused antigen with high specificity and high sensitivity which was used to detect the tuberculosis.Methods The dominant epitopes of four antigens of Mycobacterium tuberculosis,MPT64,Mtb8,Mth8.4 and Mtbl6.3, were analyzed using the BIOSUN software.The gene sequence of each dominant antigen peptide was cloned by PCR,and then ligated into expression vector pBVIL1. The genes were induced to express in E coli HB101 and the anfigenicity of each antigen was detected by indirect ELISA. Results Each dominant antigen peptide and the fused antigen were expressed highly in E coli HB101. The fused antigen presented more favourable antigenicity than that of each dominant antigen peptide. Conclusion The fused antigen, 8.4- 64 - 16.3- 8,might serve as a serediagnostic candidate antigen and was used to detect the tubeulosis,

关 键 词：结核分枝杆菌 MPT64 Mtb8 MTB8.4 Mtb16.3 

分 类 号：R392.11[医药卫生—免疫学]