阪崎克罗诺杆菌ATCC 29544免疫原性蛋白的鉴定

Identification of Immunogenic Proteins in Cronobacter sakazakii ATCC29544

摘要:

克罗诺杆菌是一种新型的食源性致病菌。迄今为止，对阪崎克罗诺杆菌的免疫原和潜在毒力因子了解很少，而免疫原性蛋白在细菌的致病性方面发挥着重要的作用。本研究利用免疫蛋白组学的方法，鉴定了阪崎克罗诺杆菌ATCC 29544的免疫原性蛋白。利用固定后的阪崎克罗诺杆菌ATCC 29544菌体免疫新西兰兔，制备多克隆抗体；采用蔗糖密度梯度离心法，提取蛋白质，并利用双向电泳对蛋白质进行分离；以制备的多克隆抗体作为一抗进行免疫印迹分析，对具有免疫反应的蛋白点进行质谱鉴定，结果共鉴定出7种具有明显免疫反应的蛋白质。

Abstract:

Cronobacter spp. are emerging opportunistic pathogens. So far, little is known about the species’ immunogens and potential virulence factors. Immunogenic proteins play an important role in the pathogenicity of bacteria. In this study, the immunogenic proteins of C. sakazakii ATCC 29544 was identified using immunoproteomics. The polyclonal antibody was prepared using the formaldehyde-fixed C. sakazakii ATCC 29544 as antigen. Then the proteins of C. sakazakii ATCC 29544 were extracted using sucrose density gradient centrifugation and separated using two-dimensional electrophoresis. Western blot was used to detect the surface-exposed proteins and mass spectroscopy was used to identify them. 7 immunoreactive proteins were identified in this study.