多杀性巴氏杆菌是危害多种畜禽和野生动物的重要病原并可造成人类发病。OmpH(包括OmpH1和OmpH2)是巴氏杆菌细胞壁中的一种主要外膜蛋白,过去对OmpH致病作用的研究主要集中于OmpH1,而有关OmpH2的报道较少。本研究对OmpH1和OmpH2一级结构...多杀性巴氏杆菌是危害多种畜禽和野生动物的重要病原并可造成人类发病。OmpH(包括OmpH1和OmpH2)是巴氏杆菌细胞壁中的一种主要外膜蛋白,过去对OmpH致病作用的研究主要集中于OmpH1,而有关OmpH2的报道较少。本研究对OmpH1和OmpH2一级结构进行了比较,发现OmpH2一级结构上具有多个与OmpH1高度相似的局部区域。采用原核表达系统表达了rOmpH2,进而开展了Western blot、Far Western blot和招募抑制实验。OmpH2与巴氏杆菌感染康复血清发生了特异性反应。OmpH2能够结合宿主Fn和Plg,而且OmpH2多克隆抗体能够显著抑制多杀性巴氏杆菌对Fn和Plg的黏附。本研究表明OmpH2能够在多杀性巴氏杆菌结合宿主Fn和Plg的过程中发挥作用,增加了多杀性巴氏杆菌毒力因子和分子致病机制的认识,对巴氏杆菌病防控具有重要意义。展开更多
[Objective] This study aimed to predict the structure of protein OmpH from Pasteurella multocida C47-8 （PmC47-8） strain of yak. [Method] Online BLAST, signal peptide prediction, secondary structure prediction and pr...[Objective] This study aimed to predict the structure of protein OmpH from Pasteurella multocida C47-8 （PmC47-8） strain of yak. [Method] Online BLAST, signal peptide prediction, secondary structure prediction and protein characteristics of sequencing result of gene OmpH from PmC47-8 strain were analyzed. [Result] The similarities of gene OmpH from PmC47-8 with the published 81 OmpH genes were between 84% and 99%; a signal peptide was found with the cleavage sites between 20 and 21 in the polypeptide; secondary structure prediction showed that folding structure accounted for 49.8% and loop structure for 50.2%; it predicted that there were 7 O-glycosylation sites in OmpH protein with the amino acid residual sites of 2, 45, 48, 330, 716, 721, 723, respectively, and 2 N-glycosylation sites with the amino acid residual sites of 15 and 35. [Conclusion] This study lays the foundation for the study on the immunity of OmpH gene from yak.展开更多
多杀性巴氏杆菌是一种重要的多种动物共患病病原和人兽共患病病原。OmpH是多杀性巴氏杆菌的保护性抗原和宿主细胞粘附因子。很多细菌的粘附因子可以通过粘附Fn等宿主胞外基质分子或血浆纤维蛋白溶解酶原(Plg)从而促进粘附过程,OmpH可能...多杀性巴氏杆菌是一种重要的多种动物共患病病原和人兽共患病病原。OmpH是多杀性巴氏杆菌的保护性抗原和宿主细胞粘附因子。很多细菌的粘附因子可以通过粘附Fn等宿主胞外基质分子或血浆纤维蛋白溶解酶原(Plg)从而促进粘附过程,OmpH可能也具有此功能。为了研究OmpH对宿主Fn和Plg的粘附作用,采用原核表达系统表达OmpH,通过Far Western Blot试验和招募抑制试验研究了OmpH对Fn和Plg的结合作用及OmpH多克隆抗体对多杀性巴氏杆菌结合这些分子的抑制作用。结果显示,重组OmpH对Fn和Plg有结合作用,OmpH抗体能够显著抑制多杀性巴氏杆菌对Fn和Plg的粘附。本研究初步证明OmpH能够通过结合宿主Fn和Plg促进多杀性巴氏杆菌的粘附作用,增加了多杀性巴氏杆菌毒力因子和分子致病机制的认识,对巴氏杆菌病防控具有积极意义。展开更多
本试验旨在构建贝氏柯克斯体外膜蛋白H(outer membrance protein H,OmpH)的重组表达载体并分析该蛋白的免疫原性,以贝氏柯克斯体九里株为模板,通过PCR方法扩增出含798bp的贝氏柯克斯体的OmpH基因片段,并将其克隆至原核表达载体pQE-30,...本试验旨在构建贝氏柯克斯体外膜蛋白H(outer membrance protein H,OmpH)的重组表达载体并分析该蛋白的免疫原性,以贝氏柯克斯体九里株为模板,通过PCR方法扩增出含798bp的贝氏柯克斯体的OmpH基因片段,并将其克隆至原核表达载体pQE-30,得到重组表达质粒pQE-30/OmpH。经IPTG诱导后,SDS-PAGE结果发现,该重组蛋白大小约为25ku。Western blotting试验结果显示,该方法所诱导产生的重组蛋白具有良好反应原性。展开更多
文摘多杀性巴氏杆菌是危害多种畜禽和野生动物的重要病原并可造成人类发病。OmpH(包括OmpH1和OmpH2)是巴氏杆菌细胞壁中的一种主要外膜蛋白,过去对OmpH致病作用的研究主要集中于OmpH1,而有关OmpH2的报道较少。本研究对OmpH1和OmpH2一级结构进行了比较,发现OmpH2一级结构上具有多个与OmpH1高度相似的局部区域。采用原核表达系统表达了rOmpH2,进而开展了Western blot、Far Western blot和招募抑制实验。OmpH2与巴氏杆菌感染康复血清发生了特异性反应。OmpH2能够结合宿主Fn和Plg,而且OmpH2多克隆抗体能够显著抑制多杀性巴氏杆菌对Fn和Plg的黏附。本研究表明OmpH2能够在多杀性巴氏杆菌结合宿主Fn和Plg的过程中发挥作用,增加了多杀性巴氏杆菌毒力因子和分子致病机制的认识,对巴氏杆菌病防控具有重要意义。
基金Supported by the Project for High-level Talents of Qinghai University (2008-QGC-7)~~
文摘[Objective] This study aimed to predict the structure of protein OmpH from Pasteurella multocida C47-8 （PmC47-8） strain of yak. [Method] Online BLAST, signal peptide prediction, secondary structure prediction and protein characteristics of sequencing result of gene OmpH from PmC47-8 strain were analyzed. [Result] The similarities of gene OmpH from PmC47-8 with the published 81 OmpH genes were between 84% and 99%; a signal peptide was found with the cleavage sites between 20 and 21 in the polypeptide; secondary structure prediction showed that folding structure accounted for 49.8% and loop structure for 50.2%; it predicted that there were 7 O-glycosylation sites in OmpH protein with the amino acid residual sites of 2, 45, 48, 330, 716, 721, 723, respectively, and 2 N-glycosylation sites with the amino acid residual sites of 15 and 35. [Conclusion] This study lays the foundation for the study on the immunity of OmpH gene from yak.
文摘多杀性巴氏杆菌是一种重要的多种动物共患病病原和人兽共患病病原。OmpH是多杀性巴氏杆菌的保护性抗原和宿主细胞粘附因子。很多细菌的粘附因子可以通过粘附Fn等宿主胞外基质分子或血浆纤维蛋白溶解酶原(Plg)从而促进粘附过程,OmpH可能也具有此功能。为了研究OmpH对宿主Fn和Plg的粘附作用,采用原核表达系统表达OmpH,通过Far Western Blot试验和招募抑制试验研究了OmpH对Fn和Plg的结合作用及OmpH多克隆抗体对多杀性巴氏杆菌结合这些分子的抑制作用。结果显示,重组OmpH对Fn和Plg有结合作用,OmpH抗体能够显著抑制多杀性巴氏杆菌对Fn和Plg的粘附。本研究初步证明OmpH能够通过结合宿主Fn和Plg促进多杀性巴氏杆菌的粘附作用,增加了多杀性巴氏杆菌毒力因子和分子致病机制的认识,对巴氏杆菌病防控具有积极意义。
文摘本试验旨在构建贝氏柯克斯体外膜蛋白H(outer membrance protein H,OmpH)的重组表达载体并分析该蛋白的免疫原性,以贝氏柯克斯体九里株为模板,通过PCR方法扩增出含798bp的贝氏柯克斯体的OmpH基因片段,并将其克隆至原核表达载体pQE-30,得到重组表达质粒pQE-30/OmpH。经IPTG诱导后,SDS-PAGE结果发现,该重组蛋白大小约为25ku。Western blotting试验结果显示,该方法所诱导产生的重组蛋白具有良好反应原性。
