两种微孢子虫孢子表面蛋白及对家蚕侵染性的比较研究  

Comparative Study on Spore Surface Proteins of Two Microsporidia and Their Infectivity to Silkworm Bombyx mori

作　　者：黄少康[1,2] 鲁兴萌[1] 汪方炜[1] 金伟[1] 陈盛禄[1] 

机构地区：[1]浙江大学动物科学学院无脊椎动物病理实验室,杭州310029 [2]福建农林大学蜂学学院

出　　处：《蚕业科学》2004年第2期157-163,共7页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目 (编号 3 0 2 70 898)

摘　　要：对来自家蚕的内网虫属样微孢子虫和家蚕微孢子虫的形态、表面蛋白及侵染性进行了比较研究。内网虫属样微孢子虫孢子显著小于家蚕微孢子虫 ,孢囊中孢子数为 8～ 32个 ,只侵染中肠组织 ,对家蚕无胚胎传染性。SDS UreaPAGE和 2D PAGE图谱显示两种孢子表面蛋白有明显差异。在SDS UreaPAGE中 ,内网虫属样微孢子虫的主要表面蛋白为 2 2kD ,而家蚕微孢子虫为 4 5kD。 2D PAGE显示 ,当上样量为 12 0 μg时 ,内网虫属样微孢子虫孢子表面蛋白斑点有 330多个 ,而家蚕微孢子虫孢子只有 16 0多个 ,其中仅有 10多个相同或疑似蛋白点。家蚕微孢子虫的主要表面蛋白点约有 2 0个 ,以中性偏酸的小分子居多 (<18kD ,pI 5 .4～ 7.2 ) ;内网虫属样微孢子虫有 30多个 ,分布较分散。根据两种孢子都能感染家蚕的特点 ,认为相同或疑似蛋白与是否能够感染有关 ,而大量差异蛋白与对家蚕的感染程度有关。两孢子表面蛋白中的主要蛋白均为差异蛋白 ,可能是孢子表面的结构蛋白。Comparative study on morphology, spore surface proteins and infectivity of a Endoreticulatus-like microsporidium and Nosema bombycis from silkworm, Bombyx mori, were conducted. The Endoreticulatus-like microsporidian spore size was significantly smaller than that of N.bombycis, and the number of spores in the sporophorous vesicle of the former ranged from 8 to 32, and midgut was the only tissue infected without transovarial transmission. The SDS-Urea PAGE and 2D-PAGE patterns of both the spore surface proteins were remarkably different. The molecular weight of main protein band of Endoreticulatus-like microsporidium was 22 kD in SDS-Urea PAGE gel, and that of N. bombycis was 45 kD. When 120 μg surface protein was loaded in 2D-PAGE gel, the Endoreticulatus-like microsporidian spore revealed over 330 protein spots, while only around 160 protein spots of N. bombycis could be observed. Comparative analysis exhibited that over 10 protein spots were suspected to be in the same position of both samples. Main protein spots of N. bombycis were around 20, mostly with low molecular weight and neutral to acidic pH (<18 kD, pI 5.4～7.2), but in Endoreticulatus-like microsporidium about 30 main protein spots that were scattered. Since both microsporidia were infective to silkworms, our analysis suggest that the proteins which were suspected to be same were responsible for their infective nature to the host, while the differential proteins maybe related to the degree of infection. The main differential surface proteins may be the structural proteins.

关 键 词：表面蛋白 侵染性 内网虫属样微孢子虫 家蚕微孢子 双向电泳 

分 类 号：S88421[农业科学—特种经济动物饲养]