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牛分枝杆菌“19 kDa”抗原基因的克隆与特性分析

Cloning and characterization of the gene for the '19 kDa' antigen of Mycobacterium bovis.

作者信息

Collins M E, Patki A, Wall S, Nolan A, Goodger J, Woodward M J, Dale J W

机构信息

Department of Microbiology, University of Surrey, Guildford, UK.

出版信息

J Gen Microbiol. 1990 Jul;136(7):1429-36. doi: 10.1099/00221287-136-7-1429.

Abstract

Monoclonal antibody CMA134.1 reacted with a protein antigen of apparent molecular mass 22 kDa from Mycobacterium bovis and Mycobacterium tuberculosis, and with an apparently 24 kDa antigen of Mycobacterium kansasii, but not with other mycobacteria or related species. This antibody was used to screen a gene library of M. bovis in lambda gt11 and identified a recombinant clone that expressed a protein with an apparent molecular mass of 19-20 kDa. Gene expression occurred from the lac promoter in lambda gt11, but used an unidentified vector promoter, possibly that of the replication primer RNA, in the final plasmid construct. The sequence of an 840 bp fragment was determined and shown to code for a product of 15 kDa. This sequence is identical to that, independently determined, of a gene from M. tuberculosis, usually referred to as the 19 kDa antigen. The reasons for the apparent size discrepancies are discussed.

摘要

单克隆抗体CMA134.1可与来自牛分枝杆菌和结核分枝杆菌的表观分子量为22 kDa的蛋白质抗原发生反应,也可与堪萨斯分枝杆菌的一种表观分子量为24 kDa的抗原发生反应,但不与其他分枝杆菌或相关菌种发生反应。该抗体用于筛选λgt11载体中的牛分枝杆菌基因文库,并鉴定出一个表达表观分子量为19 - 20 kDa蛋白质的重组克隆。基因表达在λgt11中由lac启动子驱动,但在最终的质粒构建体中使用了一个未确定的载体启动子,可能是复制引物RNA的启动子。测定了一个840 bp片段的序列,结果显示其编码一个15 kDa的产物。该序列与独立测定的结核分枝杆菌的一个基因序列相同,该基因通常被称为19 kDa抗原。文中讨论了表观大小差异的原因。

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