细粒棘球绦虫成虫蛋白的提取及特异性成分分析

Extraction of proteins from Echinococcus granulosus adult worms and their specific components analysis

旨在获得特异性蛋白提取效果好的细粒棘球绦虫(Eg)成虫蛋白提取方法,采用7种方法提取Eg成虫蛋白,以ELISA、SDS-PAGE、Western-blot等检测方法比较各蛋白提取方法的提取效果,对7种蛋白提取物进行凝胶层析获得不同蛋白组分,经过ELISA、SDS-PAGE、Western-blot等检测各组分中特异性蛋白的含量,判断各提取方法的提取效果。结果表明,7种方法均能用于提取特异性蛋白;各蛋白提取物的ELISA、SDS-PAGE和Western-blot检测显示,PBS超声破碎蛋白中特异性蛋白含量较其他蛋白提取物高。凝胶层析和ELISA检测结果表明特异性蛋白分子质量较大。比较特异性组分占总蛋白的百分比发现,PBS液氮研磨蛋白、Tris-HCl超声破碎蛋白和PBS超声破碎蛋白中特异性组分较多。本研究通过分析结合物理化学7种方法提取蛋白,为寻找蛋白质组学标准物质提供了参考依据。

The aim of the study was to obtain a good extraction method for Echinococcus granulosus(Eg)adult protein with specific protein extraction effect,the proteins were extracted by seven methods including proteomics and their antigenicities were identified by ELISA and Western-blot for obtaining antigenecity antigens.The experimental results showed that all seven methods can be used to extract specific proteins;the ELISA,SDS-PAGE and Western-blot tests of each protein extract showed that the specific protein content in PBS ultrasonic protein was higher than those of other protein extracts.The results of gel chromatography and ELISA showed that the molecular weight of the specific protein was relatively large.Comparing the percentage of specific components to total protein,it was found that there were more specific components in PBS liquid nitrogen grinding protein,Tris-HCl ultrasonic protein and PBS ultrasonic protein.This study analyzed seven protein extraction methods to provide reference for the follow-up proteomics to find standard materials.