D-arabinose synthesis in TB using Azorhizobium as a tool

使用固氮根瘤菌作为工具合成结核病中的 D-阿拉伯糖

• 批准号: 6589461
• 负责人: Michael R McNeil
• 金额: $ 3.95万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-02-15 至 2006-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6589461
• 关键词: Escherichia coli; Europe; Mycobacterium tuberculosis; antitubercular agents; arabinose; bacterial genetics; biotechnology; carbohydrate biosynthesis; carbohydrate sequence; functional /structural genomics; gene complementation; genetic mapping; gram negative bacteria; mass spectrometry; opportunistic infections; protein structure function; regulatory gene; thin layer chromatography

DESCRIPTION (provided by applicant): Tuberculosis is a major opportunistic disease of AIDS which exacerbates the course of the illness. Furthermore, tuberculosis threatens AIDS caretakers and contacts and breeds drugs resistant strains of M. tuberculosis. Hence, new drugs, specific for M. tuberculosis are needed to help control the AIDS epidemic. D-arabinose formation is an ideal TB drug target as it is essential and specific for M. tuberculosis and D-arabinose is not found in humans. The pathway for formation of D-arabinose in M. tuberculosis has been determined by the PI; however, the genes have not been successfully identified. Thus, the purpose of this AIDS-FIRCA research is to identify the M. tuberculosis genes responsible for the synthesis of D-arabinose and determine the function of the encoded proteins. The breakthrough that has made both this research and the collaboration possible is the finding by the foreign collaborator, Dr. Holsters, of four genes in Azorhizobium caulinodans that synthesize D-arabinose (A. caulinodans is one of the very few other organisms in nature that synthesize D-arabinose besides mycobacteria). Furthermore, she discovered the existence of genes in M. tuberculosis homologous to three of the A. caulinodans genes. Therefore, we propose to identify the D-arabinose synthetic genes in M. tuberculosis by complementing specific gene knockout mutants of A. caulinodans with M. tuberculosis gene candidates. Further we propose to determine the function of each gene product by the identification of D-arabinose formation precursors in these specific gene knockout strains of A. caulinodans and by expression and enzymatic assay of the M. tuberculosis and A. caulinodans genes in E. coli. The genetics on A. caulinodans and the searches for D-arabinose precursors will be mostly performed at the foreign site while needed radioactive precursors and biochemical techniques and genetics in mycobacteria will come from the American site.

描述（由申请人提供）：结核病是加剧疾病进程的艾滋病的主要机会疾病。此外，结核病威胁着艾滋病的看护人，并培养了结核分枝杆菌的抗药性菌株。因此，需要针对结核分枝杆菌的新药来帮助控制艾滋病流行。 D-阿拉伯糖的形成是理想的结核病药物靶标，因为它对于结核分枝杆菌是必不可少的，并且在人类中找不到D-阿拉伯糖。在结核分枝杆菌中形成D-阿拉伯糖的途径已由PI确定。但是，这些基因尚未成功鉴定。因此，该AIDS-FIRCA研究的目的是确定负责合成D-阿拉伯糖的结核分枝杆菌基因并确定编码蛋白的功能。使这项研究和合作成为可能的突破是外国合作者Holsters博士的发现，该基因在Caulinodans中的四个基因合成了D-阿拉伯糖（A. caulinodans是自然界中少数其他几个生物体之一，它们合成了d- arabinose mycobacteria）。此外，她发现了与三个Caulinodans基因同源的结核分枝杆菌中存在基因。因此，我们建议通过与结核分枝杆菌基因候选物的特定基因基因敲除突变体补充结核分枝杆菌中的D-阿拉伯糖合成基因。此外，我们建议通过在这些特定的caulinodans的这些特定基因敲除菌株以及大肠杆菌中的结核分枝杆菌和蛋白曲霉基因的表达和酶促测定中鉴定D-阿拉伯糖形成前体的功能。曲霉曲霉的遗传学以及对D-阿拉伯糖前体的搜索将主要在外国遗址进行，同时需要放射性的前体和分枝杆菌中的生化技术和遗传学。