含有bla_(OXA-48)基因BL21(DE3)菌株的转录组测序与分析  

Transcriptome sequencing and analysis of BL21(DE3)strain containing bla_(OXA-48) gene

作　　者：潘亚菲 康宇婷 刘文淼 贾伟 李刚 PAN Yafei;KANG Yuting;LIU Wenmiao;JIA Wei;LI Gang(Medical Experimental Center of General Hospital of Ningxia Medical University,Yinchuan 750004,China;Ningxia Key Laboratory of Clinical Pathogenic Microbiology;Ningxia Medical Laboratory Clinical Research Center)

机构地区：[1]宁夏医科大学总医院医学实验中心,宁夏银川750004 [2]宁夏临床病原微生物重点实验室 [3]宁夏医学检验临床研究中心

出　　处：《中国病原生物学杂志》2023年第8期917-921,926,共6页Journal of Pathogen Biology

基　　金：宁夏自然科学基金项目(No.2021AAC05020);宁夏回族自治区重点研发项目(No.2021BEG03090);宁夏医科大学总医院“新入职硕士培养”项目。

摘　　要：目的了解含有bla_(OXA-48)基因的BL21(DE3)菌株其细菌转录谱的变化,探究bla_(OXA-48)基因的引入对BL21(DE3)菌株代谢及耐药性的影响。方法采用高通量转录组测序的RNA-seq技术对pET32a(+)-OXA-48-BL21(DE3)与pET32a(+)-BL21(DE3)及OXA-48菌株进行检测,测序数据处理后获得表达差异基因,差异表达基因的筛选阈值为:|log2(FoldChange)|>1且qvalue<0.005。对差异表达基因进行聚类分析,利用GO数据库和KEGG数据库对表达差异基因进行注释与富集性分析,筛选部分差异表达关键基因进行分类整理。结果pET32a(+)-OXA-48-BL21(DE3)与pET32a(+)-BL21(DE3)相比有1201个差异表达基因,其中上调592个,下调609个;GO分析表明,代谢过程、生物合成过程、分子功能、催化活性、离子结合等途径共同参与OXA-48菌株的生物过程。KEGG富集分析表明差异基因主要与碳代谢、不同环境中的微生物代谢、抗生素的生物合成、嘌呤代谢、次生代谢物的生物合成等有关。对差异表达关键基因进行整理分析,与耐药性、生物膜形成、及脂多糖合成代谢相关的差异基因有acrA、acrB、ompF、pedR、mcbR和LptD等。结论bla_(OXA-48)在BL21(DE3)菌株中的表达能调控多种与细菌耐药性及代谢相关基因的表达,这为研究耐药基因对于肠杆菌科细菌产生的影响奠定了基础,可为OXA-48菌株的流行病学监测及其感染的预防控制提供参考。Objective To investigate the changes of bacterial transcription profile of BL21(DE3)strain containing bla_(OXA-48) gene,and to explore the effects of bla_(OXA-48) gene introduction on the metabolism and drug resistance of BL21(DE3)strain.Methods The high throughput transcriptome sequencing RNA seq technology was used to detect pET32a(+)-OXA-48-BL21(DE3),pET32a(+)-BL21(DE3)and strains.After sequencing data processing,differential expression genes were obtained.The screening threshold of differential expression genes was as follows:|log2(Fold-Change)|>1 and qvalue<0.005.After cluster analysis of differentially expressed genes,GO and KEGG databases were used for annotation and enrichment analysis of differentially expressed genes,and some key differentially expressed genes were selected for classification.Results There were 1201 differentially expressed genes in pET32a(+)-OXA-48-BL21(DE3)compared with pET32a(+)-BL21(DE3),of which 592 were up-regulated and 609 were down-regulated.GO analysis showed that metabolic process,biosynthesis process,molecular function,catalytic activity,ion binding and other pathways were involved in the biological process of OXA-48 strain.KEGG enrichment analysis showed that differential genes were mainly related to carbon metabolism,microbial metabolism in different environments,antibiotic biosynthesis,purine metabolism,biosynthesis of secondary metabolites and so on.After sorting out the key differentially expressed genes,we found that acrA,acrB,ompF,pedR,mcbR and LptD were related to drug resistance,biofilm formation and lipopolysis.Conclusion The expression of bla_(OXA-48) in BL21(DE3)strain can regulate the expression of multiple genes related to bacterial resistance and metabolism.Providing a foundation for studying the impact of drug resistance genes on Enter obacteriaceae bacteria,and provides a new reference for epidemiological monitoring and infection prevention and control of OXA-48 strain.

关 键 词：碳青霉烯类肠杆菌科细菌 细菌耐药性 转录组 

分 类 号：R378[医药卫生—病原生物学]