中国农业科学Issue(21)：4534-4541,8.DOI:10.3864/j.issn.0578-1752.2013.21.016
球孢白僵菌HsbA蛋白的原核表达及免疫定位
Prokaryocyte Expression and Immune Localization of HsbA in Beauveria bassiana
摘要
Abstract
[Objective]The objective of this study is to clarify the pathogenic process of hydrophobic surface binding protein A (HsbA) in Beauveria bassiana which is an important fungus used for agricultural pests control.[Method] Cloning, subcloning and prokaryotic expression were conducted in HsbA. Polyclonal anti-peptide antibody of HsbA was generated according to the purified protein, tested using Western blot, and then used for immune-localization of HsbA protein under transmission electron microscope (TEM). [Result]HsbA fusion protein was successfully expressed in pET-30a, and the polyclonal antibody specifically bound purified HsbA protein in a Western blot experiment. The immune-localization demonstrated that HsbA protein was randomly distributed in both spore and hypha. The concentration of HsbA protein in hypha was significantly greater than that in spores. The concentration of HsbA protein in spores that in the pathogenic process was significantly higher than in spores that under normal condition, while the concentration of HsbA protein in hypha under either conditions was very high. In addition, at the infected host insect (Frankiniella occidentalis Pergande), HsbA protein was mainly located in the cuticle, where the spores infected the insect body.[Conclusion]HsbA was efficiently expressed in prokartotic system and also the polyclonal anti-peptide antibody of HsbA had good immunogenicity. In addition, the results of immune localization that HsbA is related to hypha growth. Also, during the pathogenesis, it may promote the spores’ infection at the cuticle of host insects.关键词
球孢白僵菌/HsbA蛋白/原核表达/多克隆抗体/免疫电镜Key words
Beauveria bassiana/HsbA protein/prokaryocyte expression/polyclonal antibody/immuno-electron microscope引用本文复制引用
张烨,雷仲仁,王海鸿,吉青战..球孢白僵菌HsbA蛋白的原核表达及免疫定位[J].中国农业科学,2013,(21):4534-4541,8.基金项目
国家现代农业科技产业培育项目（Z121100001212006）、现代农业产业技术体系专项资金 （）
