Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (21): 4534-4541.doi: 10.3864/j.issn.0578-1752.2013.21.016

• PLANT PROTECTION • Previous Articles     Next Articles

Prokaryocyte Expression and Immune Localization of HsbA in Beauveria bassiana

 ZHANG  Ye-1, LEI  Zhong-Ren-1, WANG  Hai-Hong-1, JI  Qing-Zhan-12   

  1. 1.State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193
    2.College of Biological Science and Engineering, Beifang University of Nationalities, Yinchuan 750021
  • Received:2013-04-15 Online:2013-11-01 Published:2013-05-13

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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.

Key words: Beauveria bassiana , HsbA protein , prokaryocyte expression , polyclonal antibody , immuno-electron microscope

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