[Objective] This study aimed to investigate the outer membrane protein （OMP） patterns of Escherichia coli 038, 053 and 075 isolates from chickens. [Method] Eight pathogenic E. coil isolates with various serotypes we...[Objective] This study aimed to investigate the outer membrane protein （OMP） patterns of Escherichia coli 038, 053 and 075 isolates from chickens. [Method] Eight pathogenic E. coil isolates with various serotypes were used as experimental materials to extract OMP by using supersonic schizolysis method and Sarcosyl. After SDS-PAGE electrophoresis, OMP patterns of the extracted products were determined based on the OMP model diagram. [Result] OMP of eight E. coil isolates with three serotypes were divided into three patterns, to be specific, 2 075 isolates respectively belonged to OMP-I and OMP-II pattern, 1 053 isolate belonged to OMP-II pattern, and 5 038 isolates belonged to OMP-I and OMP-III pattern. [Conclusion] Experimental results showed that E. coli isolates with the same serotype may belong to completely different OMP patterns, while serologically unrelated isolates may belong to the same OMP pattern. OMP of E. coil isolates with the same serotype may generate genetic differentiation; in addition, OMP of E. coli isolates with different serotypes may have different genetic correlation.展开更多
The biological effect of cerium nitrate on the outer membrane(OM) of Escherichia coli(E.coli) cell was studied,and the antim-icrobial mechanism of rare earth elements was explored.The antimicrobial effect of cerium ni...The biological effect of cerium nitrate on the outer membrane(OM) of Escherichia coli(E.coli) cell was studied,and the antim-icrobial mechanism of rare earth elements was explored.The antimicrobial effect of cerium nitrate on E.coli cell was valued by plate count method,and the morphology change of E.coli cell was observed with scanning electron microscopy(SEM) and transmission electron microscopy(TEM).The results showed that the E.coli cell suspension was flocculated when the concentration of Ce(NO3)3?6H2O...展开更多
Aeromonas hydrophila, a gram negative bacterium is a major fish pathogen and causes major economic losses to aquaculture industry. Outer membrane proteins play a significant role in its survival during different envir...Aeromonas hydrophila, a gram negative bacterium is a major fish pathogen and causes major economic losses to aquaculture industry. Outer membrane proteins play a significant role in its survival during different environmental conditions and bacterial pathogenesis. The outer membrane protein R (OmpR) is a member of the two-component regulatory system of Aeromonas hydrophila which differentially regulates the expression of OmpF or OmpC depending on the osmolarity conditions. Role of OmpR has been demonstrated in its virulence in other infectious bacteria and it is found to be a potential drug target/vaccine candidate. However, the OmpR of A. hydrophila has not been characterized. In the present study, we report recombinant expression, purification of the OmpR of A. hydrophila strain Ah17 in salt inducible E. coli GJ1158 cells. Leaky expression of rOmpR was confirmed by Western blot analysis using anti-6 × His antibody. The histidine tagged recombinant OmpR (rOmpR) (~29 kDa) was purified using Ni-NTA affinity chromatography from the soluble fraction of induced E. coli cells. The rOmpR was found to be highly immunogenic with end point titres of greater than 1:80,000. The anti-rOmpR antisera were capable of agglutinating live A. hydrophila cells, thus showing vaccine potential of the rOmpR.展开更多
Objective To evaluate the role of outer membrane protein (Omp) F-deficiency and active efflux in the accumulation of hydrophilic fluoroquinolones ciprofloxacin (CPLX) and lomefloxacin (LMLX) in resistant E. coli ...Objective To evaluate the role of outer membrane protein (Omp) F-deficiency and active efflux in the accumulation of hydrophilic fluoroquinolones ciprofloxacin (CPLX) and lomefloxacin (LMLX) in resistant E. coli strains. Methods Fluoroquinolone accumulation in bacteria and the effect of active efflux were measured by a fluorescence method. The outer membrane proteins of the bacteria were analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). E. coli strains in this study included control strains JF701 and JF703 that are OmpC- or OmpF-deficient mutants of E. coli K-12, respectively, and the fluoroquinolone susceptible strain the fluoroquinolone susceptible strain of Escherichia coli (Ecs) and its in vitroselected resistant strains R2 and R256, and the clinical resistant isolates R5 and R6. Results The steady-state accumulation concentration of each drug in Ecs appeared to be the same as in JF701, while in the OmpF- deficient strain JF703, it was 1/5 CPLX or 1/2 LMLX lower than that in JF701, but JF703 was still susceptible to fluoroquinolones. On the other hand, compared with susceptible strains, a 2- to 10-fold decrease in the accumulation of each drug was found in the resistant strains except R2, in which the accumulation was slightly higher than in JF703. After the addition of 2,4-dinitrophenol (DNP), accumulation of each drug increased, especially in resistant strains, indicating that the function of the active efflux (pump) system in these bacteria had been enhanced dramatically. Furthermore, both OmpF and OmpC in Ecs, OmpF-deficiency in R2 and R256 and OmpC-deficiency in R5 and R6 were observed.Conclusion The decreased accumulation of hydrophilic fluoroquinolones in E. coli involved OmpF-deficiency and active efflux (pump), and the latter may be an important factor.展开更多
目的了解临床分离大肠埃希菌对临床常用抗菌药物的耐药情况,并探讨其外膜蛋白与耐药性的相关性。方法收集2013年全年宁夏医科大学总医院及其分院患者标本中分离的大肠埃希菌共1058株,统计分析其标本来源和科室构成,纸片扩散法测定6种常...目的了解临床分离大肠埃希菌对临床常用抗菌药物的耐药情况,并探讨其外膜蛋白与耐药性的相关性。方法收集2013年全年宁夏医科大学总医院及其分院患者标本中分离的大肠埃希菌共1058株,统计分析其标本来源和科室构成,纸片扩散法测定6种常用抗菌药物(头孢曲松、氨曲南、左氧氟沙星、环丙沙星、庆大霉素、阿米卡星)的药敏性,荧光定量PCR法检测Omp A、Omp F、Omp C m RNA表达水平,蛋白垂直电泳检测Omp A、Omp F和Omp C蛋白的表达水平。结果 1058株大肠埃希菌耐药情况为头孢曲松(67%)、环丙沙星(65.9%)、左氧氟沙星(61.9%)、庆大霉素(50.3%)、氨曲南(48.1%)、阿米卡星(3.7%)、单药耐药(31.0%)、双药耐药(30.9%)和多重耐药(27.9%),在多重耐药株中呈现Omp C m RNA和Omp C蛋白的高表达。结论大肠埃希菌外膜蛋白Omp C表达的强弱与细菌耐药有关,可能是导致细菌耐药的原因之一。展开更多
基金Supported by China Postdoctoral Science Foundation(20100470565)Science and Technology Support Program of Hebei Province(10960408D)+1 种基金Project of Science and technology Bureau of Shijiazhuang(1150093A)Science and Technology Development Project of Qinhuangdao City(201101A182)~~
文摘[Objective] This study aimed to investigate the outer membrane protein （OMP） patterns of Escherichia coli 038, 053 and 075 isolates from chickens. [Method] Eight pathogenic E. coil isolates with various serotypes were used as experimental materials to extract OMP by using supersonic schizolysis method and Sarcosyl. After SDS-PAGE electrophoresis, OMP patterns of the extracted products were determined based on the OMP model diagram. [Result] OMP of eight E. coil isolates with three serotypes were divided into three patterns, to be specific, 2 075 isolates respectively belonged to OMP-I and OMP-II pattern, 1 053 isolate belonged to OMP-II pattern, and 5 038 isolates belonged to OMP-I and OMP-III pattern. [Conclusion] Experimental results showed that E. coli isolates with the same serotype may belong to completely different OMP patterns, while serologically unrelated isolates may belong to the same OMP pattern. OMP of E. coil isolates with the same serotype may generate genetic differentiation; in addition, OMP of E. coli isolates with different serotypes may have different genetic correlation.
基金supported by the National Natural Science Foundation of China (20971028)the Foundation of Enterprise-University-Research Institute Cooperation from Guangdong Province and Ministry of Education of China (2007B090400105,2008A010500005)the Foundation of Science and Technology Project of Yuexiu District of Guangzhou City of China (2008-PT-006)
文摘The biological effect of cerium nitrate on the outer membrane(OM) of Escherichia coli(E.coli) cell was studied,and the antim-icrobial mechanism of rare earth elements was explored.The antimicrobial effect of cerium nitrate on E.coli cell was valued by plate count method,and the morphology change of E.coli cell was observed with scanning electron microscopy(SEM) and transmission electron microscopy(TEM).The results showed that the E.coli cell suspension was flocculated when the concentration of Ce(NO3)3?6H2O...
文摘Aeromonas hydrophila, a gram negative bacterium is a major fish pathogen and causes major economic losses to aquaculture industry. Outer membrane proteins play a significant role in its survival during different environmental conditions and bacterial pathogenesis. The outer membrane protein R (OmpR) is a member of the two-component regulatory system of Aeromonas hydrophila which differentially regulates the expression of OmpF or OmpC depending on the osmolarity conditions. Role of OmpR has been demonstrated in its virulence in other infectious bacteria and it is found to be a potential drug target/vaccine candidate. However, the OmpR of A. hydrophila has not been characterized. In the present study, we report recombinant expression, purification of the OmpR of A. hydrophila strain Ah17 in salt inducible E. coli GJ1158 cells. Leaky expression of rOmpR was confirmed by Western blot analysis using anti-6 × His antibody. The histidine tagged recombinant OmpR (rOmpR) (~29 kDa) was purified using Ni-NTA affinity chromatography from the soluble fraction of induced E. coli cells. The rOmpR was found to be highly immunogenic with end point titres of greater than 1:80,000. The anti-rOmpR antisera were capable of agglutinating live A. hydrophila cells, thus showing vaccine potential of the rOmpR.
文摘Objective To evaluate the role of outer membrane protein (Omp) F-deficiency and active efflux in the accumulation of hydrophilic fluoroquinolones ciprofloxacin (CPLX) and lomefloxacin (LMLX) in resistant E. coli strains. Methods Fluoroquinolone accumulation in bacteria and the effect of active efflux were measured by a fluorescence method. The outer membrane proteins of the bacteria were analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). E. coli strains in this study included control strains JF701 and JF703 that are OmpC- or OmpF-deficient mutants of E. coli K-12, respectively, and the fluoroquinolone susceptible strain the fluoroquinolone susceptible strain of Escherichia coli (Ecs) and its in vitroselected resistant strains R2 and R256, and the clinical resistant isolates R5 and R6. Results The steady-state accumulation concentration of each drug in Ecs appeared to be the same as in JF701, while in the OmpF- deficient strain JF703, it was 1/5 CPLX or 1/2 LMLX lower than that in JF701, but JF703 was still susceptible to fluoroquinolones. On the other hand, compared with susceptible strains, a 2- to 10-fold decrease in the accumulation of each drug was found in the resistant strains except R2, in which the accumulation was slightly higher than in JF703. After the addition of 2,4-dinitrophenol (DNP), accumulation of each drug increased, especially in resistant strains, indicating that the function of the active efflux (pump) system in these bacteria had been enhanced dramatically. Furthermore, both OmpF and OmpC in Ecs, OmpF-deficiency in R2 and R256 and OmpC-deficiency in R5 and R6 were observed.Conclusion The decreased accumulation of hydrophilic fluoroquinolones in E. coli involved OmpF-deficiency and active efflux (pump), and the latter may be an important factor.
文摘目的了解临床分离大肠埃希菌对临床常用抗菌药物的耐药情况,并探讨其外膜蛋白与耐药性的相关性。方法收集2013年全年宁夏医科大学总医院及其分院患者标本中分离的大肠埃希菌共1058株,统计分析其标本来源和科室构成,纸片扩散法测定6种常用抗菌药物(头孢曲松、氨曲南、左氧氟沙星、环丙沙星、庆大霉素、阿米卡星)的药敏性,荧光定量PCR法检测Omp A、Omp F、Omp C m RNA表达水平,蛋白垂直电泳检测Omp A、Omp F和Omp C蛋白的表达水平。结果 1058株大肠埃希菌耐药情况为头孢曲松(67%)、环丙沙星(65.9%)、左氧氟沙星(61.9%)、庆大霉素(50.3%)、氨曲南(48.1%)、阿米卡星(3.7%)、单药耐药(31.0%)、双药耐药(30.9%)和多重耐药(27.9%),在多重耐药株中呈现Omp C m RNA和Omp C蛋白的高表达。结论大肠埃希菌外膜蛋白Omp C表达的强弱与细菌耐药有关,可能是导致细菌耐药的原因之一。
