本文選題：噬菌體 + 高通量測(cè)序��； 參考：《中國(guó)抗生素雜志》2017年10期

【摘要】：噬菌體是解決致病菌多重耐藥性問(wèn)題的最佳選擇之一,然而噬菌體的宿主特異性使得噬菌體的裂解譜有限,因此研究噬菌體宿主特異性對(duì)于噬菌體治療應(yīng)用具有重要意義。本研究通過(guò)篩選大腸埃希菌宿主菌BL21噬菌體IME253的耐受菌,結(jié)合對(duì)敏感菌和耐受菌的高通量測(cè)序,分析基因組差異,預(yù)測(cè)噬菌體耐受相關(guān)基因,發(fā)現(xiàn)耐受與外膜受體蛋白FepA有關(guān)。然后利用向?qū)RISPR/Cas9切開(kāi)敏感菌fepA基因,同時(shí)利用Red同源重組系統(tǒng)無(wú)痕敲除fepA基因,證明fepA基因敲除可以導(dǎo)致細(xì)菌BL21對(duì)噬菌體IME253耐受。本實(shí)驗(yàn)利用比較基因組學(xué)分析平臺(tái),建立了一種宿主耐受噬菌體的分析方法,并且利用CRIPSR無(wú)痕敲除技術(shù)來(lái)驗(yàn)證耐受相關(guān)基因,為噬菌體治療提供理論基礎(chǔ)。
[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.
【作者單位】： 河北師范大學(xué)生命科學(xué)院;青島農(nóng)業(yè)大學(xué)動(dòng)物科技學(xué)院;軍事醫(yī)學(xué)科學(xué)院微生物流行病研究所病原微生物生物安全國(guó)家重點(diǎn)實(shí)驗(yàn)室;
【基金】：科技重大專(zhuān)項(xiàng)“十二五”實(shí)施計(jì)劃項(xiàng)目(No.2013ZX10004-605、No.2013ZX10004-217、No.2013ZX10004-607、No.2011ZX10004-001) 國(guó)家高技術(shù)研究發(fā)展計(jì)劃項(xiàng)目(863計(jì)劃)(No.2014AA021402和No.2012AA022-003) 國(guó)家自然科學(xué)基金項(xiàng)目(No.81572045)
【分類(lèi)號(hào)】：Q933

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本文編號(hào)：2024506