亚洲玉米螟幼虫血浆蛋白提取方法的比较  

A comparison of protein extraction methods suitable for plasma protein of Asian corn borer larva

作　　者：王滔[1] 杨巽[1] 衣建坤[1] 王玉[1] 孙宇[1] 席景会[1] 

机构地区：[1]吉林大学植物科学学院,长春130062

出　　处：《环境昆虫学报》2013年第2期264-268,共5页Journal of Environmental Entomology

基　　金：公益性行业(农业)科研专项(201003025);吉林大学研究生创新基金资助项目(20121134)

摘　　要：为了建立适合利用双向电泳分析的亚洲玉米螟5龄幼虫血浆蛋白样品的制备平台,为后续用双向电泳技术研究不同条件下亚洲玉米螟5龄幼虫血浆蛋白的差异表达奠定基础,本研究比较了直接提取法、TCA/丙酮沉淀法、PEG提取法三种蛋白质提取方法。结果表明:直接提取法SDS-PAGE图谱中条带最少,只有22条,双向电泳图谱中的蛋白质点数较少,为332个;TCA/丙酮沉淀法同样是SDS-PAGE图谱条带较少,为23条,且双向电泳图谱中蛋白点数最少,只有210个点;而PEG提取法SDS-PAGE图谱条带数最多,为39条,且分布均匀,双向电泳图谱蛋白点数最多,为874个点,且分布广泛。通过以上结果可以得出,PEG提取法最适合用于亚洲玉米螟血浆蛋白样品的制备。To establish a suitable preparation platform of the Asian corn borer during fifth instar stage plasma protein sample for 2 - DE analysis and provide a platform for the Asian corn borer during fifth instar stage plasma protein sample＇s preparation using Two - Dimensional Eleetrophoresis （2 - DE） , we can lay the foundation for the differential expression of the plasma protein in different conditions using 2 - DE in the follow -up research and experiments. This study compared three protein extraction methods： Direct extraction method, TCA/acetone precipitation and PEG extraction method. The results indicated that： Direct extraction method analyzed by SDS -PAGE electrophoresis showed the least, only 22 bands and protein spots in 2 - DE Map were less, 332 ; similarly, TCA/acetone precipitation method have just 23 bands analyzed by SDS - PAGE electrophoresis and protein spots in 2 - DE Map were the least, 210 ; The PEG extraction method showed the most, 39 protein bands evenly distributed in SDS -PAGE map, protein spots widely distributed in 2 - DE map were the most, 874. These results displayed that PEG extraction method was the most appropriate method for the preparation of the Asian corn borer plasma protein.

关 键 词：亚洲玉米螟 血浆蛋白 提取方法 双向电泳 

分 类 号：Q966[生物学—昆虫学] S435.13[农业科学—农业昆虫与害虫防治]