Abstract:

Abstract: 【OBJECTIVE】In order to detect expression of α-Mannosidase and study its possible function during infection of Magnaporthe oryzae, a polyclonal antibody of a putative α-Mannosidase was prepared. 【METHOD】 A transformant of Magnaporthe oryzae, overexpressing a putative α-Mannosidase-His tag fusion protein was grown in liquid media. The α-Mannosidase fusion protein was purified by Ni2+ -NTA affinity chromatography column from culture filtrate. The fusion protein was then injected into and used to immune New Zealand rabbits to produce polyclonal antibody. 【RESULTS】The titer of this polyclonal antibody was 1：51, 200 and with good specificity after ELISA analysis. Protein of 30 M. oryzae strains were tested against this antibody using ELISA and Western blotting show that the expression level of the α- Mannosidase appeared to be significantly different among strains as well as infection stages when inoculated with the strain 70-15. 【CONCLUSION】The difference expression level of the α- Mannosidase among different strains may be related to the physiological and biological characteristic such as pathogenicity of the fungus.