Abou-Zeid C, Garbe T, Lathigra R, Wiker H G, Harboe M, Rook G A, Young D B
Department of Medical Microbiology, University College and Middlesex School of Medicine, London, United Kingdom.
Infect Immun. 1991 Aug;59(8):2712-8. doi: 10.1128/iai.59.8.2712-2718.1991.
Recombinant phage clones, TB1 and TB2, were selected from a Mycobacterium tuberculosis lambda gt11 DNA expression library by screening with a polyclonal antiserum raised against the antigen 85 complex of Mycobacterium bovis BCG. Analysis of recombinant DNA inserts and expressed fusion proteins showed that two new genes had been isolated. The product of clone TB2 was identified as a member of the 30/31-kDa antigen 85 complex. Restriction enzyme analysis showed that this gene differs from previously cloned members of this antigen complex, with detailed serological analysis indicating that it may encode the 85C component. Antisera raised against the expressed product of clone TB1 recognized a 55-kDa protein in M. tuberculosis extracts. The 55-kDa protein also has fibronectin-binding activity and, like the 30/31-kDa family, is a prominent target of the antibody response in patients with mycobacterial disease. Although the clones were selected by using the same antiserum, detailed analysis by serology and by DNA hybridization showed that they represent two quite distinct types of fibronectin-binding activities expressed by M. tuberculosis. Further analysis of the fibronectin-binding antigens of M. tuberculosis may provide important insights into their role in mediating the interaction with the host immune system.
通过用针对卡介苗牛分枝杆菌抗原85复合物产生的多克隆抗血清进行筛选，从结核分枝杆菌λgt11 DNA表达文库中选出了重组噬菌体克隆TB1和TB2。对重组DNA插入片段和表达的融合蛋白的分析表明分离出了两个新基因。克隆TB2的产物被鉴定为30/31-kDa抗原85复合物的一个成员。限制性酶切分析表明该基因与该抗原复合物先前克隆的成员不同，详细的血清学分析表明它可能编码85C成分。针对克隆TB1表达产物产生的抗血清识别结核分枝杆菌提取物中的一种55-kDa蛋白。该55-kDa蛋白也具有纤连蛋白结合活性，并且与30/31-kDa家族一样，是分枝杆菌病患者抗体反应的主要靶标。尽管这些克隆是用相同的抗血清筛选出来的，但血清学和DNA杂交的详细分析表明它们代表了结核分枝杆菌表达的两种截然不同类型的纤连蛋白结合活性。对结核分枝杆菌纤连蛋白结合抗原的进一步分析可能为深入了解它们在介导与宿主免疫系统相互作用中的作用提供重要线索。
