荔枝胚蛋白质的提取方法

以不同体积的Tris-HCl(0.1mol／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(Lithi chinensis)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。

A Method for Extracting Embryo Proteins in Litchi chinensis Sonn.

Different volumes of Tris-HCl buffer(0.1mol/L,pH8.8)in combination with different contents of PVP40were employed to study the embryo protein extracted from Litchi chinensis varieties"Huaizhi ","Guiwei"and"Heiye".It was shown that the volume of Tris-HCl buffer being5times the fresh wieght of embryo and added with15%PVP40(g g -1 FW)was adapted to obtain a satisfactory result.Homogenate containing15%PVP40(g g -1 FW)could be used directly for determination of soluble protein content by Coomassie Brilliant Blue G-250and10%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 containing2%PVP40.Precipitation of embryo and endosperm proteins with ethanol of10times the volume of protein extraction gave best SDS-PAGE pattern.