人血小板生成素受体c-MPL膜内部分的聚合作用  

Aggregation of the Intracellular Domain of the Human Thrombopoietin Receptor c MPL

作　　者：赵新燕[1] 王光清[1] 周伟国 陈春妹 李昌本[1] 赵寿元[1] 

机构地区：[1]复旦大学遗传学研究所遗传工程国家重点实验室

出　　处：《生物化学与生物物理学报》1998年第6期565-569,共5页

基　　金：国家自然科学基金;国家"863"计划生物技术领域青年基金

摘　　要：血小板生成素（ｔｈｒｏｍｂｏｐｏｉｅｔｉｎ，ＴＰＯ）是调节血小板生成最主要的细胞因子，其生物学效应由其受体ｃ－ＭＰＬ介导。利用酵母双杂合（ｔｗｏ－ｈｙｂｒｉｄ）系统研究ｃ－ＭＰＬ膜内部分在ＴＰＯ信号转导途径中的功能。首先用反转录ＰＣＲ（ＲＴ－ＰＣＲ）方法从人红白血病ＨＥＬ细胞系总ＲＮＡ中扩增并克隆Ｐ型ｃ－ＭＰＬ（ＭＰＬＰ）膜内部分ｃＤＮＡ，经测序验证后克隆至双杂合载体ｐＡＳ２和ｐＧＡＤ４２４中，重组质粒命名为ｐＡＳＭＭ和ｐＧＡＤＭＭ。将ｐＡＳＭＭ与ｐＧＡＤＭＭ质粒共转化酵母菌，可激活报告基因ｈｉｓ３和ｌａｃＺ的表达，从而证明ＭＰＬＰ膜内部分在体内可以相互作用，这种聚合作用在ＴＰＯ信号转导途径中起重要作用。Thrombopoietin (TPO) is the major cytokine which is involved in platelet production and exerts its effects via the receptor c MPL. The yeast two hybrid system has been used to study the aggregation of the intracellular domain of c MPL in TPO signal transduction. First, the cDNA fragment of MPLP intracellular domain was amplified and cloned by using RT PCR method from the total RNA of HEL cells. Then the cDNA fragment was sequenced and subcloned into two hybrid vectors pAS2 and pGAD424, respectively, and the recombinants are named as pASMM and pGADMM. Co transformation of these plasmids into yeast activated his 3 and lac Z reporter genes, demonstrating in vivo interaction of the MPLP receptor intracellular domain itself. The aggregation may be important in TPO signal transduction.

关 键 词：血小板生成素 酵母双杂合系统 聚合作用 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]