Phenotypically profiling a Mycobacterium bovis mutant library, through a chemical-genetic screening

通过化学遗传筛选对牛分枝杆菌突变体文库进行表型分析

• 批准号: 2266926
• 金额: --
• 依托单位: University of Birmingham
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2266926
• 关键词: Phenotypically; profiling; Mycobacterium; bovis; mutant

Mycobacterium tuberculosis and M. bovis BCG can adapt to various chemical and environmental stresses and thereby evade therapeutic treatment. The genetic basis for this response remains mysterious, as up to 40% of the genome is unannotated and functional classification of many important genes remains elusive. High-throughput reverse chemical genetics have proven immensely powerful in dissecting and annotating the gene-network of complex cellular processes. I will use this technology to gain a detailed, genome-wide stress-response map of the tuberculosis vaccine strain BCG. I will subject a transposon derived, single gene deletion library (~10,000 mutants) to ~300 mycobacterial drugs and environmental perturbations to mimic the harsh environments M. tuberculosis and BCG can encounter. This will generate a comprehensive fitness signature for each mutant and perturbation tested, allowing me to: (i) discover functions for orphan genes, (ii) identify pathways and/or processes involved in antimicrobial resistance and (iii) record drug and mutant responses to identify primary and secondary targets. We envision that those insights may improve antimicrobial therapies for humans and livestock.

结核分枝杆菌和牛分枝杆菌能适应各种化学和环境胁迫，从而逃避治疗。这种反应的遗传基础仍然是神秘的，因为高达40%的基因组是未注释的，许多重要基因的功能分类仍然难以捉摸。高通量反向化学遗传学在解剖和注释复杂细胞过程的基因网络方面被证明是非常强大的。我将使用这项技术获得结核疫苗株卡介苗的详细的全基因组应激反应图。我将把转座子衍生的单基因缺失文库（约10,000个突变体）置于约300种分枝杆菌药物和环境扰动中，以模拟结核分枝杆菌和卡介苗可能遇到的恶劣环境。这将为测试的每个突变和扰动生成一个全面的适应度签名，使我能够：(i)发现孤儿基因的功能，（ii）确定与抗菌素耐药性有关的途径和/或过程，（iii）记录药物和突变反应，以确定主要和次要目标。我们设想这些见解可能会改善人类和牲畜的抗菌疗法。