2株毒力不同M.bovis差异表达蛋白的定量检测及分析  

Quantitative Determination and Analysis of Differentially Expressed Proteins in Two Strains of Mycobacterium bovis with Different Virulence

作　　者：李梦莹 田帅[1] 董玉惠 马小静 张淮瑜 宋阿北 周向梅[2] 许立华[1] LI Meng-ying;TIAN Shuai;DONG Yu-hui;MA Xiao-jing;ZHANG Huai-yu;SONG A-bei;ZHOU Xiang-mei;XU Li-hua(Agriculture College,Ningxia University,Yinchuan 750021,China;College of Veterinary Medicine,China Agricultural University,Beijing 100193,China)

机构地区：[1]宁夏大学农学院,宁夏银川750021 [2]中国农业大学动物医学院,北京海淀100193

出　　处：《中国兽医杂志》2021年第1期6-9,14,I0004,I0005,共7页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金项目(31560687,31760722,2017YFD0500901);宁夏大学研究生创新项目(GIP2019006)。

摘　　要：为研究牛分枝杆菌N和C68004菌株间蛋白水平的差异表达对其致病性的影响,本试验采用串联质谱标记(TMT)定量蛋白组学技术,对2个菌株的毒力及其致病性进行鉴定与分析。结果分析得到2174个共有蛋白,其中有479个差异表达蛋白(P<0.05)。GO分析结果显示,表达差异显著蛋白的功能包括催化、结合、运输、转录调控以及分子结构相关作用,特别是对内部或外部刺激产生的反应。KEGG分析结果显示,表达差异显著蛋白主要参与代谢和生物过程的调节。导致N菌致病性增强的毒力基因可能是mmaA 4、ecce 1、IpqY、hspX、Mpb 63和mmpl 4。并且N菌的耐药基因rpoA、rpoB和rpoC表达量显著高于C68004,可以推断N菌对抗结核一线药物利福平的耐药性可能比C68004更强。随着临床用药频繁和免疫应答的改变,N菌在适应环境过程中,与代谢、DNA复制以及耐药相关的蛋白表达出现了差异。In order to study the influence of different protein expression levels on the pathogenicity of Mycobacterium bovis N and C68004 strains,this study used tandem mass tag(TMT)spectrometry for quantitative proteomics technology to identify and analyze the virulence and pathogenicity of the two strains.Results showed that there were 2174 common proteins and 479 differentially expressed proteins with P<0.05.GO analysis results showed that the functions of the differentially expressed proteins included catalysis,binding,transport,transcriptional regulation and molecular structure-related roles,especially in response to internal or external stimulation.KEGG analysis results showed that the differentially expressed proteins were mainly involved in the regulation of metabolic and biological processes.The virulence genes that caused increased pathogenicity in M.bovis N might be mmaA 4,ecce 1,IpqY,hspX,Mpb 63,and mmpl 4.In addition,the expression levels of rpoA,rpoB and rpoC of N were significantly higher than that of C68004,suggesting that the resistance of N to rifampicin,the first-line tuberculosis drug,is stronger than that of C68004.With frequent clinical medication and changes in immune response,the expression of proteins related to metabolic,DNA replication and drug resistance of N was different in the process of adaptation to the environment.

关 键 词：牛分枝杆菌 蛋白组学 差异表达蛋白 毒力差异 

分 类 号：Q939.131[生物学—微生物学]