禽巴氏杆菌环丙沙星耐药株转录组测序与分析  

The transcriptome sequencing analysis of ciprofloxacin-resistant avian Pasteurella multocida

作　　者：孔令严[1,2] 罗青平[2] 卢琴[2] 邵华斌[2] 李槿年[1] 

机构地区：[1]安徽农业大学动物科技学院,安徽合肥230036 [2]湖北省农业科学院畜牧兽医研究所,湖北武汉430064

出　　处：《中国预防兽医学报》2017年第4期262-266,共5页Chinese Journal of Preventive Veterinary Medicine

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)

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。To identify the differentially expressed genes of sensitive strains and ciprofloxacin （Cip） resistant strains of avian Pasteurella multocida, the transcriptomes were sequenced by high-throughput RNA sequencing. All of the genes were obtained from avian P.multocida （Pm70） control strain （NC_002663.1） using its reference genome to analyze the yield and quality of assembly, which were annotated using the BLAST search against GO and KEGG databases. The differentially expressed genes were enriched in gene ontology and KEGG pathway, also were verified byreal-time PCR. The results showed that the mapping ratios of reads were above 95.87% of all the samples; A total of 19 differential expressing genes were identified including 15 up-regulated genes and 4 down-regulated genes in Cip-resistant strain. The GO analyses that differential expression genes clustered significantly was transporter activity. The KEGG analyses that the enrichment pathways that are most represented was ＂ABC transporters＂ （ko02010）. It suggested that ABC transporters might play important roles in the mechanisms of Cip-resistance in avian P.multocida.

关 键 词：禽源多杀性巴氏杆菌 转录组测序 GO功能分析 KEGG功能分析 耐药机制 

分 类 号：S852.61[农业科学—基础兽医学]