摘要
为建立适于SDS-PAGE分析的蚜虫蛋白质样品制备平台,以便为蚜虫蛋白质的双向电泳分析奠定基础,本研究比较了TCA/丙酮沉淀、PEG提取、饱和酚抽提和直接裂解4种蛋白质提取方法。结果表明:不同样品制备方法的蛋白提取率有显著的差异,其中直接裂解法的提取率最高,为17.43mg/g;其次是饱和酚抽提法,提取率为12.30mg/g;而PEG制备法提取率最低,只有7.96mg/g。利用SDS-PAGE电泳对不同的蛋白质样品进行了分析,发现在凝胶图谱上显现的条带也有明显的差异,其中饱和酚抽提法显现的条带数最多,为36条,且从14.4kDa～116.0kDa范围有广泛分布,条带清晰;PEG提取法条带数为30条,一些蛋白条带丢失或不明显;TCA/丙酮沉淀法的蛋白条带集中分布在25.0kDa～67.0kDa区域;直接裂解法条带数仅为24条,且小分子量的条带可辩率很低。通过以上结果可以得出,饱和酚抽提法最适用于蚜虫全蛋白样品的制备。
To establish a suitable preparation platform of aphid protein sample for SDS-PAGE analysis, we compared four protein extraction methods： TCA/acetone precipitation, PEG, saturated phenol, and direct decomposition method. The results indicated that, extraction rate of different protein preparation methods were significant different by statistical analysis. The direct decomposition method presented the highest yield of 17.43 mg/g, followed by the saturated phenol extraction, 12.30 mg/g. The yield of PEG extraction method was the lowest, only 7.96 mg/g. There was also an obvious difference in protein bands of samples from different protein preparation methods, analyzed by SDS-PAGE electrophoresis. The saturated phenol extraction method showed most, 36 bands, broadly and clearly distributing from 14.4 kDa to 116.0 kDa range. The PEG extraction method showed 30 protein bands, a number of bands was lost or became unapparent. Protein bands of TCA/acetone precipitation method concentrated in the 25.0 kDa-67.0 kDa region. The direct decomposition method was only 24 bands, and the bands of small molecular weight protein with low resolution. These results displayed that the saturated phenol extraction was the most appropriate method for aphid protein preparation for SDS-PAGE analysis.
出处
《环境昆虫学报》
CSCD
北大核心
2010年第3期353-356,共4页
Journal of Environmental Entomology
基金
国家高技术研究发展计划(2008AA10Z224)
吉林省科技厅基础研究项目(20060545)
关键词
蚜虫
蛋白提取
电泳
aphid
protein extraction
SDS-PAGE
