荔枝胚蛋白质的提取方法

A Method for Extracting Embryo Proteins in Litchi chinensis Sonn.

摘要:

以不同体积的Tris-HCl(0.1 mol/L,pH8.8)为提取液,结合不同含量(以胚鲜重计)的PVP40,对怀枝、黑叶和桂味等荔枝(Litchichinensis)品种的胚蛋白质进行提取。结果表明,提取液体积为胚鲜重的5倍(ml g^-1 FW),并加入15%的PVP40时,提取蛋白质的效果最好,可用于荔枝胚可溶性蛋白质含量的测定;胚乳蛋白质的提取则以等体积的提取液(内含2%的PVP40)为佳。加入10%PVP40的胚蛋白提取液可直接进行SDS-PAGE电泳,用10倍于蛋白质提取液体积的乙醇沉淀胚和胚乳的蛋白提取液,可得到最佳的SDS-PAGE电泳效果。

Abstract:

Different volumes of Tris-HCl buffer (0.1 mol/L, pH 8.8) in combination with different contents of PVP40 were employed to study the embryo protein extracted from Litchi chinensis varieties "Huaizhi", "Guiwei" and "Heiye". It was shown that the volume of Tris-HC1 buffer being 5 times the fresh wieght of embryo and added with 15% PVP40 (g g^-1 FW) was adapted to obtain a satisfactory result. Homogenate containing 15% PVP40 (g g^-1 FW) could be used directly for determination of soluble protein content by Coomassie Brilliant Blue G-250 and 10% PVP40 (g g^-1 FW) for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Endosperm protein could be extracted with equal volume of Tris-HCl containing 2% PVP40. Precipitation of embryo and endosperm proteins with ethanol of 10 times the volume of protein extraction gave best SDS-PAGE pattern.