水稻根部蛋白质组样品制备的方法研究

为了选择适用于水稻根部蛋白质组分析的样品制备方法,以水稻苗期幼嫩根尖为材料,采用TCA/丙酮沉淀法、Tris法、尿素/硫脲法对水稻根部总蛋白质进行提取并比对,通过Bradford法、SDS PAGE电泳技术对蛋白质含量和种类进行分析。结果显示,Tris法提取出的水稻根部蛋白质含量和种类明显少于其他2种方法;尿素/硫脲法和TCA/丙酮沉淀法提取的蛋白质种类和含量都较多,其中,尿素/硫脲法的蛋白质图谱中蛋白条带多,染色深而且很清晰,尤其适用于较低分子量蛋白质的提取,TCA/丙酮沉淀法则适于高分子量蛋白质的提取。因此,进行水稻根部总蛋白质组分析时,可选用尿素/硫脲法。

A Study on Sample Preparation Method of Rice Root Proteome

In order to develop the sample preparation methods for rice root proteome analysis,the TCA/acetone precipitation method,direct extraction method and urea/thiourea method were used to extract the rice root protein,and the protein contents and types of samples were analyzed using Bradford method and SDS-PAGE electrophoresis.The results indicated that the direct extraction method was the worst among the three methods,giving the least protein bands and protein content;the TCA/acetone precipitation method and the urea/thiourea method were the optimal methods,both producing more protein bands,while the latter showed more protein types and more clear bands,especially for the low-molecular weight proteins,while the TCA/acetone precipitation method was more suitable for the preparation of high molecular weight proteins.It was suggested that different methods had different specialties and the urea/thiourea method was the optimal method for total protein extraction of rice roots.