| 摘要: |
| 【目的】比较鼠伤寒沙门菌标准株及其环丙沙星体外诱导耐药株菌的蛋白表达图谱差异，分析这些蛋白在鼠伤寒沙门菌诱导耐药过程中的作用，为沙门菌耐药机理的研究提供参考。【方法】将沙门菌标准株和环丙沙星耐药株在LB培养基中培养至对数期，然后超声破碎细菌制备蛋白样品，进行二维电泳，电泳胶经扫描分析后选取差异表达蛋白点进行质谱鉴定。【结果】沙门菌环丙沙星耐药株与标准株有25个差异表达蛋白，其中磷酸化孔蛋白PhoE、二氢硫辛酰胺脱氢酶组件蛋白、ATP合成酶、细胞侵袭蛋白SipD的A亚基、锰过氧化氢酶、二氢硫辛酰胺转琥珀酰酶、果糖二磷酸醛缩酶、假定蛋白AF355_20900等8个蛋白表达量下调，磷酸乙酰基转移酶、天冬氨酸转氨酶、RND家族外排泵的膜融合蛋白、鞭毛马达开关FliM、三磷酸甘油醛脱氢酶、OmpA外膜蛋白、蛋白质链伸长因子EF Ts、丙二醇利用蛋白PduB、NAD(P)H硝基还原酶、假定硫醇 烷基过氧化氢还原酶、50S核糖体亚基L4、肽ABC转运体结合蛋白、假定的胞质蛋白、锰超氧化物歧化酶、30S核糖体S4亚基、TolC外膜蛋白、短链脱氢酶等17个蛋白表达量上调。这25个差异蛋白中，有1个（1号）与抗药性无关，有2个（8和21号）为未知功能蛋白，其余22个是与细菌耐药性相关的蛋白。这22个蛋白可分为4类，其中耐药相关蛋白5个、毒力相关蛋白2个、蛋白质合成相关蛋白3个、代谢相关蛋白12个；其亚细胞定位为：细胞质中的蛋白15个、外膜蛋白3个、细胞质内膜蛋白2个、周质蛋白和细胞外蛋白各1个。【结论】与沙门菌标准株相比，耐环丙沙星沙门菌有25个差异表达蛋白（8个表达下调，17个表达上调），其中22个与耐药性相关，分属4个功能类别，定位于5个亚细胞组分。 |
| 关键词: 鼠伤寒沙门菌 耐药性 环丙沙星 体外诱导 蛋白表达图谱 |
| DOI： |
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| 基金项目:国家自然科学基金项目（31772802）；安徽高校自然科学重点研究项目（KJ2017A132）；安徽省优秀青年人才支持计划重点项目（gxyqZD2016041） |
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| Difference in protein expression between ciprofloxacin resistant strain and standard strain of Salmonella |
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ZHANG Ruiliang,XU Jun,WANG Rui,et al
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| Abstract: |
| 【Objective】This study compared bacterial protein expression profiles of Salmonella typhimurium standard strain and ciprofloxacin strain in vitro and analyzed the role of proteins in induction of drug resistance to provide reference for understanding drug resistance mechanism of Salmonella.【Method】Salmonella standard strains and ciprofloxacin resistant strains were cultured in logarithmic phase in LB medium.Protein samples were prepared by ultrasonic disruption of bacteria,and two dimensional electrophoresis was carried out before the protein spots in electrophoretic gel were selected for mass spectrometry identification.【Result】 There were 25 differentially expressed proteins between Salmonella ciprofloxacin resistant strains and standard strains,among which phosphorylated porin PhoE,dihydrolipoamide dehydrogenase component protein,ATP synthase,cell invasion protein SipD,manganese catalase,dihydrolipoamide amide succinylase,fructose diphosphate aldolase,and putative protein AF355_20900 had down regulated expression,while phosphor acetyltransferase,aspartate aminotransferase,RND membrane fusion protein of family efflux pump, flagellar motor switch FliM,glyceraldehyde triphosphate dehydrogenase,OmpA outer membrane protein, protein chain elongation factor EF-Ts,propylene glycol utilization protein PduB,NAD(P)H nitroreductase,putative thiol alkyl hydroperoxide reductase,50S ribosomal subunit protein L4,peptide ABC transporter binding protein,putative cytoplasmic protein, manganese superoxide dismutase,30S ribosomal S4 subunit,TolC outer membrane protein,short-chain dehydrogenase had up-regulated expression.Of the 25 differential proteins,1 (No. 1) was not related to drug resistance,2 (8 and 21) were unknown functional proteins,and the remaining 22 were bacterial resistance related proteins.These 22 proteins can be divided into 4 categories, including 5 resistance related proteins,2 virulence-related proteins,3 protein-related proteins,and 12 metabolic related proteins.The subcellular localization showed there were 15 proteins in the cytoplasm,3 outer membrane proteins,2 cytoplasmic inner membrane proteins,1 periplasmic protein,and 1 extracellular protein.【Conclusion】There were 25 differentially expressed proteins between Salmonella ciprofloxacin strain (8 down regulated and 17 up-regulated) and standard strain,22 of which were related to drug resistance,belonging to 4 functional categories and located in 5 subcellular fractions. |
| Key words: Salmonella drug resistance ciprofloxacin induction in vitro protein expression prafile |
