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

结核分枝杆菌和M.牛BCG可以适应各种化学和环境应激，从而逃避治疗性治疗。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已被证明是非常强大的解剖和注释复杂的细胞过程的基因网络。我将利用这项技术来获得结核病疫苗株BCG的详细的全基因组应激反应图谱。我将转座子衍生的单基因缺失文库（约10，000个突变体）应用于约300种分枝杆菌药物和环境扰动以模拟恶劣环境M。结核病和卡介苗可能会遇到。这将为测试的每个突变体和扰动生成全面的适应性特征，使我能够：（i）发现孤儿基因的功能，（ii）识别抗菌素耐药性中涉及的途径和/或过程，以及（iii）记录药物和突变体反应以识别主要和次要靶标。我们设想这些见解可以改善人类和牲畜的抗菌治疗。

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.