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

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.