禽巴氏杆菌四环素耐药株转录组测序与分析
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摘要
为鉴定禽源多杀性巴氏杆菌(avian Pasteurella multocida,Pm)四环素(Tetracycline,Tet)敏感株与耐药株(MIC=32μg/mL)在转录组水平上的差异,本研究采用RNA-seq技术对Tet敏感株与耐药株进行双向测序,通过GO、KEGG和ARDB数据库对表达差异基因进行注释和富集性分析。结果显示,共筛选出442个差异表达基因,其中包括103个上调基因和339个下调基因。通过GO功能富集分析显示差异表达基因多数富集于核糖体结构成分、rRNA结合、跨膜运输和离子转运等生物过程;对差异表达基因进行信号通路富集性分析,大多数差异表达基因分布在ABC转运蛋白与核糖体代谢途径中;对差异表达基因进行耐药基因注释(ARDB),发现其中的耐药基因主要为核糖体保护蛋白和外排系统两类。本研究表明禽源Pm对四环素耐药的耐药机制可能与细胞膜上ABC转运蛋白的过量表达以及核糖体保护蛋白有关。
To identify the differentially expressed genes of Tetracycline sensitive strains and resistant strains(MIC=32μg/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(GO), Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway and Antibiotic Resistance Genes Database(ARBD), respectively. The results showed that 442 differentially expressed genes were identi?ed, including 103 up-regulated genes and 339 down-regulated genes. The GO enrichment analysis showed that the majority of differentially expressed genes play a role in "structural constituent of ribosome", "rRNA binding", "transmembrane transport" and "ion transport". Most of the differentially expressed genes were distributed in the "ABC transporters" and "Ribosome" by KEGG analysis. The differentially expressed resistance genes were mainly divided into ribosomal protection protein and ef?ux pump system by ARDB analysis. In summary,ABC transporters and aminoglycoside transferase overexpression may play important roles in the mechanisms of Tet-resistance in avian P.multocida.
To identify the differentially expressed genes of Tetracycline sensitive strains and resistant strains(MIC=32μg/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(GO), Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway and Antibiotic Resistance Genes Database(ARBD), respectively. The results showed that 442 differentially expressed genes were identi?ed, including 103 up-regulated genes and 339 down-regulated genes. The GO enrichment analysis showed that the majority of differentially expressed genes play a role in "structural constituent of ribosome", "rRNA binding", "transmembrane transport" and "ion transport". Most of the differentially expressed genes were distributed in the "ABC transporters" and "Ribosome" by KEGG analysis. The differentially expressed resistance genes were mainly divided into ribosomal protection protein and ef?ux pump system by ARDB analysis. In summary,ABC transporters and aminoglycoside transferase overexpression may play important roles in the mechanisms of Tet-resistance in avian P.multocida.
引文
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[23]Furushita M,Shiba T,Maeda T,et al.Similarity of tetracycline resistance genes isolated from fish farm bacteria to those from clinical isolates[J].Appl Environ Microbiol,2003,69(9):5336-5342.
[2]皮家贵,李雨南,万婷婷,等.禽巴氏杆菌病的诊断与防治[J].当代畜禽养殖业,2017(1):24.
[3]San Millan A,Escudero J A,Gutierrez B,et al.Multiresistance in Pasteurella multocida is mediated by coexistence of small plasmids[J].Antimicrob Agents Chemother,2009,53(8):3399-3404.
[4]孟丹,孟昱,张斌,等.耐药性禽巴氏杆菌的分离鉴定及耐药性检验[J].吉林畜牧兽医,2009,30(11):33-34.
[5]孔令聪,战利,赵晴,等.动物源多杀性巴氏杆菌耐药性及耐药机制研究进展[J].中国兽药杂志,2012,46(3):52-55.
[6]孙广龙,胡立宏.四环素类抗生素的研究进展[J].药学研究,2017,36(1):1-5.
[7]Dhamgaye S,Bernard M,Lelandais G,et al.RNAsequencing revealed novel actors of the acquisition of drug resistance in Candida albicans[J].BMC Genomics,2012,13:396.
[8]魏树全,钟维农,郑业勤,等.泛耐药铜绿假单胞菌体外耐药模型构建及其耐药机制的研究[J].中华医院感染学杂志,2012,22(13):2731-2735.
[9]王平,樊志奇,范瑞强.香莲外洗液逆转白念珠菌耐药的转录组学研究[J].中国中西医结合杂志,2015,35(12):1505-1509.
[10]Bustin S A,Benes V,Garson J A,et al.The MIQEguidelines:minimum information for publication of quantitative real-time PCR experiments[J].Clin Chem,2009,55(4):611-622.
[11]Kehrenberg C,Schwarz S.Molecular analysis of tetracycline resistance in Pasteurella aerogenes[J].Antimicro Agents Chemother,2001,45(10):2885-2890.
[12]冯新,韩文瑜,雷连成.细菌对四环素类抗生素的耐药机制研究进展[J].中国兽药杂志,2004(2):38-42.
[13]徐宗香,李阳,赵志慧,等.猪链球菌对四环素类抗生素耐药机制的研究进展[J].猪业科学,2011(2):84-87.
[14]米阳,郑鹏远,张炳勇,等.外排泵ABC转运蛋白基因msbA和spab在幽门螺杆菌多重耐药中的重要作用[J].世界华人消化杂志,2011,19(14):1500-1505.
[15]Gall A R,Datsenko K A,Figueroa-Bossi IV,et al.Mg2+regulates transcription of mgtA in Salmonella Typhimurium via translation of proline codons during synthesis of the MgtL peptide[J].Proc Natl Acad Sci USA,2016,113(52):15096-15101.
[16]Wang S,Gao X,Gao Y,et al.Tetracycline resistance genes identified from distinct soil environments in China by functional metagenomics[J].Front Microbiol,2017,8:1406.
[17]Adesoji A T,Ogunjobi A A,Olatoye I O,et al.Prevalence of tetracycline resistance genes among multi-drug resistant bacteria from selected water distribution systems in southwestern Nigeria[J].Ann Clin Microbiol and Antimicrob,2015,14:41.
[18]Dintner S,Heermann R,Fang C.A sensory complex consisting of an ATP-binding cassette transporter and a two-component regulatory system controls bacitracin resistance in Bacillus subtilis[J].J Biol Chem,2014,289(40):27899-27910.
[19]W?sten M M,van de Lest C H,van Dijk L,et al.Function and regulation of the C4-dicarboxylate transporters in Campylobacter jejuni[J].Front Microbiol,2017,8:174.
[20]Valentini M,Storelli N,Lapouge K.Identification of C4-dicarboxylate transport systems in Pseudomonas aeruginosa PAO1[J].J Bacteriol,2011,193(17):4307-4316.
[21]Xiong J,Feng J,Yuan D.Tracing the structural evolution of eukaryotic ATP binding cassette transporter superfamily[J].Sci Rep,2015,5:16724.
[22]Xu S,Fu Z,Zhou Y,et al.Mutations of the transporter proteins GlpT and UhpT confer fosfomycin resistance in Staphylococcus aureus[J].Front Microbiol,2017,8:914.
[23]Furushita M,Shiba T,Maeda T,et al.Similarity of tetracycline resistance genes isolated from fish farm bacteria to those from clinical isolates[J].Appl Environ Microbiol,2003,69(9):5336-5342.