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当前位置: 首页 > 文章 > 柞蚕微孢子虫3种孢壁蛋白的提取与鉴定 蚕业科学 2011,37 (2) 155-161
Position: Home > Articles > Extraction and Identification of Three Spore Wall Proteins from Nosema pernyi Science of Sericulture 2011,37 (2) 155-161

柞蚕微孢子虫3种孢壁蛋白的提取与鉴定

作  者：
王伯阳;姜义仁;杨瑞生;李艳卓;王勇;秦利
单  位：
沈阳农业大学生物科学技术学院
关键词：
柞蚕微孢子虫;孢壁蛋白;基质辅助激光解析电离飞行时间质谱
摘  要：
在微孢子虫侵染宿主的过程中,孢壁蛋白扮演着重要的角色。分别采用沸水浴法、SDS法从柞蚕微孢子虫(Nosema per-nyi,Np)孢子中提取孢壁蛋白以及利用发芽后的孢子壳提取孢壁蛋白,对提取的孢壁蛋白进行SDS-PAGE分析和质谱鉴定。结果显示,将发芽后的孢子壳以沸水浴法提取孢壁蛋白最为有效,但操作复杂,部分孢壁蛋白损失严重。初步鉴定从发芽后的孢子壳中提取的编号为P35、P32和P29的3种蛋白属于柞蚕微孢子虫孢壁蛋白,蛋白分子质量分别为35、32、29 kD,其中孢壁蛋白P32的含量最丰富,P35和P29的含量相对较低。根据相应肽指纹图谱预测孢壁蛋白P32的一级结构类似核孔蛋白,可能与孢壁的选择透过性有关,而P35与VASA2n蛋白的一级结构类似,因此有可能与柞蚕微孢子虫的繁殖有关。
译  名：
Extraction and Identification of Three Spore Wall Proteins from Nosema pernyi
作  者：
WANG Bo-Yang JIANG Yi-Ren YANG Rui-Sheng LI Yan-Zhuo WANG Yong QIN Li(College of Bioscience and Biotechnology,Shenyang Agricultural University,Shenyang 110866,China)
关键词：
Nosema pernyi;Spore wall protein;MALDI-TOF-Mass
摘  要：
The spore wall proteins of Nosema pernyi play an important role when it infects the host insect Antheraea pernyi.The present study extracted spore wall proteins of N.pernyi by boiling method and SDS method from N.pernyi spores and germinated spore coats,and characterized the obtained spore wall proteins by SDS-PAGE and mass spectrometry.The results showed that the extraction of spore wall proteins from spore coats by boiling method was the most efficient.But the operation was complex and some spore wall proteins were lost heavily.Three different proteins numbered P35,P32 and P29 were preliminarily identified to be spore wall proteins of N.pernyi with molecular weight of 35 kD,32 kD and 29 kD respectively.The content of P32 was the highest,while those of P35 and P29 were relatively lower.Peptide fingerprinting predicted that the primary structure of P32 was similar to nuclear pore proteins,suggesting its function related to selective permeability of spore wall,and that of P35 was similar to VASA2n,suggesting its involvement in the propagation of N.pernyi.

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