禽巴氏杆菌链霉素耐药株转录组测序与分析  

The transcriptome sequencing analysis of Streptomycin-resistant strains of avian Pasteurella multocida

作　　者：汪最 孔令严 邵华斌 卢琴 罗青平 WANG Zui;KONG Ling-yan;SHAO Hua-bin;LU Qin;LUO Qing-ping(Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan 430064, China;College of Animal Sciences, Yangtze University, Jingzhou 434025, China;Key Laboratory of animal disease control and prevention formulation of Ministry of Agriculture, Wuhan 430064, China)

机构地区：[1]湖北省农业科学院畜牧兽医研究所,湖北武汉430064 [2]长江大学动物科学学院,湖北荆州434025 [3]农业部畜禽细菌病防治制剂创制重点实验室,湖北武汉430064

出　　处：《中国预防兽医学报》2018年第4期288-293,共6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(201303040);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(Pm)链霉素(Str)敏感株与耐药株(MIC=1024μg/mL)在转录组水平上的差异,本研究采用RNA-seq技术对Str敏感株与耐药株进行双向测序,筛选得到差异表达基因,通过GO、KEGG和ARDB数据库对差异表达基因进行分组和富集性分析,并利用荧光定量PCR对筛选出的差异表达基因进行验证。结果显示,共筛选获得625个差异表达基因,其中包括581个上调基因和44个下调基因。通过GO功能富集分析显示差异表达基因多数发挥细胞膜转运功能;对差异表达基因进行信号通路富集性分析显示,大多数差异表达基因分布在ABC转运蛋白与双组分系统代谢途径中;采用ARDB对差异表达基因进行耐药基因分析,显示其中的耐药基因主要为氨基糖苷转移酶类和外排系统两类。本研究结果表明,禽源Pm对Str的耐药机制可能与细胞膜上多药外排泵的过量表达和氨基糖苷转移酶类灭活抗菌药物有关。同时,也为进一步研究禽源Pm对Str的耐药机制奠定了基础。To identify the differentially expressed genes of Streptomycin sensitive strains and resistant strains （MIC=1024μg/mL） of avian P.multocida, the transcriptomes were detected by high-throughput RNA sequencing. All of differentially expressed genes were enriched and annotated .by gene ontology and KEGG pathway, at the same time, they were verified by qRT-PCR. The results showed that 625 differentially expressed genes were identified, including 581 up-regulated genes and 44 down-regulated genes in Str-resistant strain. GO enrichment analysis showed that the majority of differentially expressed genes played a role in cell membrane transport; Most of the differentially expressed genes were distributed in the ＂ABC transporters＂ and ＂Two-component system＂ by KEGG analysis; the differentially expressed resistance genes were mainly divided into aminoglycoside transferase and efflux pump system by ARDB analysis. In summary, ABC transporters and aminoglycoside transferase overexpression might play important roles in the mechanisms of Str-resistance in avian P.multocida. This study also provides an important basis for further research on the molecular mechanisms of Str-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 ARDB分析 

分 类 号：S852.61[农业科学—基础兽医学]