禽巴氏杆菌环丙沙星耐药株转录组测序与分析

发布时间：2019-01-13 06:52

【摘要】：为鉴定禽源多杀性巴氏杆菌(Avian Pasteurella multocida,Pm)敏感株与环丙沙星(Cip)耐药株的差异表达基因,本研究采用高通量转录组测序的RNA-seq技术对敏感株与Cip耐药株进行检测,参考Pm70株基因组(NC_002663.1)进行拼接与质量评估,测序数据处理后获得表达差异基因,利用GO和KEGG数据库对表达差异基因进行注释与富集性分析,并利用荧光定量PCR对转录组测序中的差异表达基因进行验证。结果显示,所有样品测序数据参考Pm70全基因组匹配率均达到95.87%以上,对基因差异表达进行分析后鉴定得到19个差异表达基因,其中15个上调,4个下调。通过GO功能富集分析显示差异表达基因主要为转运运输功能,并对差异表达基因进行信号通路富集性分析,结果表明最具代表性的信号通路为"ABC转运蛋白"(ko02010)。推测禽源Pm中的ABC转运蛋白超家族可能在Cip耐药机制中具有重要作用。
[Abstract]:In order to identify the differentially expressed genes between susceptible strains of Pasteurella multocida (Avian Pasteurella multocida,Pm) and ciprofloxacin resistant strains (Cip), high throughput transcriptional sequencing (RNA-seq) technique was used to detect sensitive strains and Cip resistant strains. Pm70 genome (NC_002663.1) was used for splicing and quality evaluation. After sequencing data were processed, the differentially expressed genes were obtained. The GO and KEGG databases were used to annotate the differentially expressed genes and to analyze the enrichment of the differentially expressed genes. Fluorescent quantitative PCR was used to verify the differentially expressed genes in transcriptome sequencing. The results showed that the total genome matching rate of all samples was over 95.87%, and 19 differentially expressed genes were identified by analysis of gene differential expression, of which 15 were up-regulated and 4 down-regulated. GO functional enrichment analysis showed that the differentially expressed genes were mainly transporter, and the signal pathway enrichment of differentially expressed genes was analyzed. The results showed that the most representative signal pathway was "ABC transporter" (ko02010). It is suggested that the ABC transporter superfamily in avian Pm may play an important role in the mechanism of Cip resistance.
【作者单位】：安徽农业大学动物科技学院;湖北省农业科学院畜牧兽医研究所;
【基金】：公益性农业行业科研专项:家禽主要细菌病防控技术研究与示范(201303044) 现代农业产业技术体系建设专项资金资助(CARS-42-G11) 规模化养鸡场主要疫病防控关键技术研究(2015ABA039)
【分类号】：S852.61

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