CHARACTERIZATION OF MYCOBACTERIUM INTRACELLULARE LAMBDA-GT11 RECOMBINANT CLONES

在过去的一年里，分枝杆菌实验室继续进行了 遗传、生化和免疫学特征评价 两种免疫反应性胞内分枝杆菌抗原。 MI 43-核苷酸序列分析先前已经表明， MI 43基因编码27 kDa的脂蛋白。最近，我们证实 生物化学，使用去污剂相分离实验和代谢 用~ 3 H-棕榈酸标记MI 43为脂蛋白。 我们有 也证明纯化的MI 43被70%的血清识别 从结核病患者中，80%的艾滋病毒血清阴性血清 50%的MAC患者血清和50%的MAC AIDS患者血清 疾病 MI 85- Our核苷酸序列分析及在大肠杆菌中的表达 大肠杆菌的研究表明，MI 85基因编码过氧化氢酶-过氧化物酶， 双功能酶，可能是关键的吞噬体内 分枝杆菌的存活。由于军情85处的重要性，我们必须 产生了许多单克隆抗体， 这个抗原。使用这些抗体之一的诱导实验具有 证明了MI 85在体外被过氧化物诱导三倍。 在 此外，我们还鉴定了含有MI 85的核苷酸序列， 过氧化氢酶启动子，目前正在表征结构和 启动子的活性。最后，我们已经克隆并正在表征 过氧化氢酶-过氧化物酶基因在M.结核和M. avium

During the past year, the Laboratory of Mycobacteria has continued the evaluation of the genetic, biochemical, and immunologic characterization of two immunoreactive Mycobacterium intracellulare antigens. MI43 - Nucleotide sequence analyses have previously indicated that the MI43 gene encodes a 27 kDa lipoprotein. Recently, we have confirmed biochemically, using detergent phase separation experiments and metabolic labeling with 3H palmitic acid that MI43 is a lipoprotein. We have also demonstrated that purified MI43 is recognized by 70% of the sera from patients with tuberculosis, 80% of sera from HIV-seronegative patients with MAC disease, and 50% of sera from AIDS patients with MAC disease. MI85 - Our nucleotide sequence analysis and expression in Escherichia coli suggest that the MI85 gene encodes a catalase-peroxidase, a bi-functional enzyme that may be critical for the intraphagosomal survival of mycobacteria. Because of the importance of MI85, we have generated a number of monoclonal antibodies which recognize epitopes on this antigen. Induction experiments using one of these antibodies have demonstrated that MI85 is induced three-fold by peroxide in vitro. In addition, we have identified the nucleotide sequences containing the MI85 catalase promoter and are currently characterizing the structure and activity of the promoter. Finally, we have cloned and are characterizing the catalase-peroxidase genes of M. tuberculosis and M. avium.