利用比较基因组学快速鉴定大肠埃希菌噬菌体耐受基因

发布时间：2018-06-16 01:08

本文选题：噬菌体 + 高通量测序　；参考：《中国抗生素杂志》2017年10期

【摘要】：噬菌体是解决致病菌多重耐药性问题的最佳选择之一,然而噬菌体的宿主特异性使得噬菌体的裂解谱有限,因此研究噬菌体宿主特异性对于噬菌体治疗应用具有重要意义。本研究通过筛选大肠埃希菌宿主菌BL21噬菌体IME253的耐受菌,结合对敏感菌和耐受菌的高通量测序,分析基因组差异,预测噬菌体耐受相关基因,发现耐受与外膜受体蛋白FepA有关。然后利用向导CRISPR/Cas9切开敏感菌fepA基因,同时利用Red同源重组系统无痕敲除fepA基因,证明fepA基因敲除可以导致细菌BL21对噬菌体IME253耐受。本实验利用比较基因组学分析平台,建立了一种宿主耐受噬菌体的分析方法,并且利用CRIPSR无痕敲除技术来验证耐受相关基因,为噬菌体治疗提供理论基础。
[Abstract]:Bacteriophage is one of the best choice to solve the problem of multidrug resistance of pathogenic bacteria. However, the host specificity of bacteriophage makes the cleavage spectrum of phage limited, so it is important to study the specificity of phage host for the application of bacteriophage therapy. In this study, we screened the resistant bacteria of Escherichia coli BL21 phage IME253, combined with the high-throughput sequencing of sensitive bacteria and tolerant bacteria, analyzed the genomic differences, predicted the genes related to phage tolerance, and found that tolerance was related to the outer membrane receptor protein FepA. Then the fepA gene was cut open by CRISPRP / Cas9 and the fepA gene was knocked out by the Red homologous recombination system. It was proved that fepA gene knockout could induce the bacterial BL21 tolerance to the bacteriophage IME253. In this study, a host tolerance phage analysis method was established by using comparative genomics analysis platform, and CRIPSR marker free knockout technique was used to verify tolerance related genes, which provided a theoretical basis for phage therapy.
【作者单位】：河北师范大学生命科学院;青岛农业大学动物科技学院;军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室;
【基金】：科技重大专项“十二五”实施计划项目(No.2013ZX10004-605、No.2013ZX10004-217、No.2013ZX10004-607、No.2011ZX10004-001) 国家高技术研究发展计划项目(863计划)(No.2014AA021402和No.2012AA022-003) 国家自然科学基金项目(No.81572045)
【分类号】：Q933

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