Dissection of Mycobacterium tuberculosis antigens using recombinant DNA.
Dissection of Mycobacterium tuberculosis antigens using recombinant DNA.
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使用重组 DNA 解剖结核分枝杆菌抗原。
DOI：
10.1073/pnas.82.9.2583
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发表时间：
1985
影响因子：
11.1
通讯作者：
Davis,RW
中科院分区：
文献类型：
--
作者：
Young,RA;Bloom,BR;Grosskinsky,CM;Ivanyi,J;Thomas,D;Davis,RW
重组DNA策略已经被用来系统地调查结核分枝杆菌基因组，以寻找编码由单抗检测到的特定抗原的序列。利用随机产生终点的结核分枝杆菌基因组DNA片段，构建了大的lambda gt11重组DNA表达文库。产生了足够数量的重组子，以包含其端点出现在病原体基因组中几乎每个碱基对的插入物。用单抗探针筛选病原菌DNA线性片段所特有的重组噬菌体产生的蛋白抗原。这种方法与一种改进的基因分离检测方法相结合，该方法使用抗体克隆分离指定结核分枝杆菌多肽成分的DNA序列。这里描述的方法适用于其他病原体，为开发更敏感和特异的结核病免疫诊断和血清流行病学测试提供了可能性，并最终为开发更有效的疫苗提供了可能性。
A recombinant DNA strategy has been used systematically to survey the Mycobacterium tuberculosis genome for sequences that encode specific antigens detected by monoclonal antibodies. M. tuberculosis genomic DNA fragments with randomly generated endpoints were used to construct a large lambda gt11 recombinant DNA expression library. Sufficient numbers of recombinants were produced to contain inserts whose endpoints occur at nearly every base pair in the pathogen genome. Protein antigens specified by linear segments of pathogen DNA and produced by the recombinant phage of Escherichia coli were screened with monoclonal antibody probes. This approach was coupled with an improved detection method for gene isolation using antibodies to clonally isolate DNA sequences that specify polypeptide components of M. tuberculosis. The methodology described here, which is applicable to other pathogens, offers possibilities for the development of more sensitive and specific immunodiagnostic and seroepidemiological tests for tuberculosis and, ultimately, for the development of more effective vaccines.