IDENTIFICATION OF PREF 1 INTERACTING PROTEIN

本研究的长期目标是：（1）探索 在脂肪细胞的分化和分化过程中， 胚胎发生(2)探讨前脂肪细胞的分子机制 分化及其与人类疾病的联系。具体目标 该方案的主要目的是（1）鉴定与β-1相互作用的蛋白质; (2)以完成编码相互作用蛋白的cDNA。主 将努力集中在与蛋白质相互作用的蛋白质上。 胞外结构域（ECD）。将鉴别蛋白质 通过其与纯化的EST-I融合蛋白的亲和结合进行生物化学反应 和/或内源性表达的IFN-γ-1蛋白。三种常规方法， GST-下拉测定、交联测定和共- 将应用免疫沉淀分析。开放式阅读框架 编码β-1相互作用蛋白的基因将由其中一个 以下方法：（1）使用来自纯化的DNA探针， 相互作用蛋白或放射性标记的GST-GST-1（ECD）来探测cDNA 利用3 T3-L1细胞构建表达文库;（2）酵母双杂交 筛选; 3）.平移。

The long-term objectives of this research are: (1) To explore the regulatory function of pref-1 in both adipocyte differentiation and embryogenesis. (2) To pursue the molecular mechanism of preadipocyte differentiation and its connection with human disease. The specific aims of this proposal are (1) to identify the pref-1 interacting protein(s); (2) to done the cDNA encoding the interacting protein(s). The primary efforts will be focused on protein(s) that interacts with the extracellular domain (ECD) of pref-1. The protein(s) will be identified biochemically by its affinity binding to purified pref-1 fusion proteins and/or endogenous expressed pref-1 protein. Three conventional methods, the GST-pulldown assay, the crosslinking assay, and the co- immunoprecipitation assay will be applied. The open reading frame encoding the pref-1 interacting protein(s) will be doned by one of the following methods: (1) using a DNA probe derived from purified interacting protein or radiolabeled GST-pref-1(ECD) to probe a cDNA expression library made from the 3T3-L1 cells; (2) yeast two-hybrid screening; 3). panning.