Shinnick T M
J Bacteriol. 1987 Mar;169(3):1080-8. doi: 10.1128/jb.169.3.1080-1088.1987.
The immune response of the host to the antigens of Mycobacterium tuberculosis plays the key role in determining immunity from infection with as well as the pathogenicity of this organism. A 65-kilodalton (kDa) protein has been identified as one of the medically important antigens of M. tuberculosis. The gene encoding this antigen was isolated from a lambda gt11-M. tuberculosis recombinant DNA library using monoclonal antibodies directed against the 65-kDa antigen as the specific probes. The nucleotide sequence of this gene was determined, and a 540-amino-acid sequence was deduced. This sequence was shown to correspond to that of the 65-kDa antigen by constructing a plasmid in which this open reading frame was fused to the lacZ gene. The resulting fusion protein reacted specifically with the anti-65-kDa protein antibodies. A second long open reading frame was found downstream of the 65-kDa antigen gene which could encode a protein of 517 amino acids. This putative protein contained 29 tandemly arranged partial or complete matches to a pentapeptide sequence.
宿主对结核分枝杆菌抗原的免疫反应在决定对该菌感染的免疫力以及其致病性方面起着关键作用。一种65千道尔顿(kDa)的蛋白质已被确定为结核分枝杆菌的重要医学抗原之一。使用针对65-kDa抗原的单克隆抗体作为特异性探针,从λgt11-结核分枝杆菌重组DNA文库中分离出编码该抗原的基因。测定了该基因的核苷酸序列,并推导了一个540个氨基酸的序列。通过构建一个将该开放阅读框与lacZ基因融合的质粒,证明该序列与65-kDa抗原的序列相对应。所得融合蛋白与抗65-kDa蛋白抗体发生特异性反应。在65-kDa抗原基因下游发现了第二个长开放阅读框,它可以编码一个517个氨基酸的蛋白质。这种推定的蛋白质包含29个与一个五肽序列串联排列的部分或完全匹配。