志贺菌敏感株与基因转移耐多药株蛋白组学分析  

Analysis on proteomics between sensitive strain and gene transferredmulti-drug resistant strain of shigella flexneri

作　　者：宋春花[1] 黄学勇[1] 郗园林[1] 张梅喜[1] 段广才[1] 

机构地区：[1]郑州大学公共卫生学院,河南450001

出　　处：《中国公共卫生》2008年第1期42-45,共4页Chinese Journal of Public Health

基　　金：河南省科技公关项目(0424410035)

摘　　要：目的对志贺菌敏感株与基因转移耐多药株全菌蛋白进行蛋白质组比较,寻找细菌耐多药相关蛋白。方法采用接合基因转移实验对临床分离鉴定志贺菌敏感株进行基因转移耐多药试验;并对志贺菌敏感株及基因转移耐多药株全菌蛋白进行双向电泳;电泳图谱采用Image Master 2D Platinum软件分析,并对差异表达蛋白进行基质辅助激光解吸飞行时间质谱(MOLDITOF-TOF)分析。结果成功获得志贺菌基因转移耐多药株,在志贺菌敏感株与耐多药株全菌蛋白质图谱中分析别检出(946±37)和(1 013±157)个蛋白质斑点;经分析共发现43个差异表达的蛋白点,初步对其中7个差异表达蛋白进行质谱鉴定,基因转移耐多药株中发现2个新出现的耐多药相关蛋白分别为成簇插入的DNA重复序列(CRISPR)相关蛋白及Hsp60的分子伴侣蛋白(Groel-Groes-Adp7);ATP结合盒(ABC)转运蛋白、胱氨酸合成酶、预测的胞浆脂蛋白表达量上调;翻译延长因子Tu及DNA单链结合蛋白表达量下降。结论对7个耐多药相关蛋白鉴定分析表明,供体菌中一些耐多药相关基因通过基因转移方式插入志贺菌敏感株中并大量表达,同时一些在细胞代谢中起重要作用的酶类表达量上调,ABC转运蛋白在志贺菌基因转移耐多药机制中也起重要作用。Objective In order to search for the new proteins realated to multi-drug resistance,the proteomics of whole cellular proteins between sensitive strain and gene transferred multi-drug resistance strain of shigella flexneri by using of two-dimensional electrophoresis（2-DE） were compared.Methods Clinical sensitive isolates shigella flexneri was transduced to multi-durg resistance stain by conjugation experiment,immobilized pH gradient（IPG） two-dimensional electrophoresis was adopted and the gels were analyzed by Image Master 2D Platinum software,matrix assisted laser desorption/ionization-time of flight mass spectrometry（MALDI-TOF MS） was used to analyze differential expression proteins.Results Gene transferred multi-durg resistance strain of shigella flexneri was obtained successfully.It was found that there were（946±37） protein spots in whole cellular protein 2-DE gels of sensitive strain,and（1013±157） protein spots in that of gene transferred multi-durg resistance strain,43 differential expression protein spots were found and 7 proteins related to multi-drug resistance were identified based on peptide mass fingerprinting.CRISPR-associated protein and Groel-Groes-Adp7 were two novel proteins;ATP binding cassette transporter protein,cysteine synthase and predicted periplasmic or secreted lipoprotein protein were high expression in gene transferred multi-durg resistance strain;But China elongation factor EF-Tu and SSB were high expression in sensitive strain.Conclusion By mass spectrum analysis of 7 proteins related to multi-durg resistance,it was found that some genes related to multi-durg resistance of donator were transferred into sensitive stain and expressed highly as well as some important cellular metabolism enzymes.ATP binding cassette transporter protein influenced seriously in multi-durg resistant mechanism of shigella flexneri.

关 键 词：志贺菌 耐多药 蛋白质组学 双向电泳 质谱分析 差异表达 

分 类 号：R378.2[医药卫生—病原生物学]