茶苗叶片低温诱导蛋白提取分离研究

Research of Isolating Low-temperature-induced Protein from Tea Seedling by Different Extracting Methods

本文研究冷驯化过程中茶苗叶片细胞膜相对透性和可溶性蛋白含量的变化,采用不同方法提取分离冷驯化茶苗叶片的低温诱导蛋白。结果表明:①与离体叶片相比,冷驯化茶苗叶片更适宜用来分离茶树低温诱导蛋白,其表现为质膜相对透性变化不大,而且可溶性蛋白含量保持在较高水平。②与丙酮沉淀法、TCA/丙酮沉淀法、硫酸铵沉淀法和直接提取法相比,改良Tris-HCl法最适于冷驯化茶苗叶片低温诱导蛋白的分离提取,其特点是蛋白样品得率高、蛋白条带多且清晰、杂质干扰程度少、操作步骤简易。另外,在冷驯化蛋白提取样品中还发现了大小分别为25、35和90 KDa的蛋白差异条带。

The effects of low temperature on cell memberane penetrability and soluble protein content in tea seedling leaves were studied.Different methods were used to extract and isolate low-temperature-induced proteins from tea seedling leaves after cold acclimation treatment.The results showed that tea seedling leaves treated by cold acclimation was propitious to induce low-temperature-induced proteins compared with the abscised leaves.Cell memberane penetrability of those seedling leaves were tested to have a tiny change,and the soluble protein content maintained a certain level.Compared with acetone precipitation method,TCA/ acetone precipitation method,ammonium sulphate precipitation method and direct extraction method,improved Tris-HCl extraction methods was the optimum method for extracting low-temperature-induced proteins in tea seedling leaves.This method showed the characteristics of high protein yields,clear protein band,few impurity disturbance and easy operation.In addition,in the SDS-PAGE electrophoretic pattern,three new protein bands（25,35,90 KDa） of the extraction samples appeared.