粘膜系统鳞状细胞癌(SCC)相关蛋白(DCUN1D1)的表达、纯化和晶体生长  

Expression,Purification and Crystallization of DCUN1D1(SCC-related oncogene)in Mucosal System

作　　者：杨晓瑜[1] 谷欣[2] 郭满才[1] 刘迎芳[2] 

机构地区：[1]西北农林科技大学,杨凌712100 [2]中国科学院生物物理研究所,北京100101

出　　处：《现代生物医学进展》2007年第7期1002-1004,共3页Progress in Modern Biomedicine

摘　　要：目的:SCCRO/RP42/DCUN1D1是粘膜系统鳞片状细胞癌(SCC)发生时人类基因组3q区域扩增的潜在靶标之一,其蛋白作用机制尚不清楚,本文拟通过表达并大量纯化SCC相关蛋白DCUN1D1用于蛋白结晶以求获得其三维结构。方法:使用人肝脑组织RNA反转录产物为模板扩增出DCUN1D1基因cDNA片断并将其克隆至原核表达载体PGEX-6P-1中,通过IPTG诱导获得大量可溶性表达,再经过GST亲和层析和Sephadex G-200层析柱纯化。结果:获得了纯度95%以上的蛋白,采用悬滴气相扩散法筛选蛋白晶体,获得显微镜下可见的微晶。结论:初步得出DCUN1D1晶体生长条件及范围,为解析DCUN1D1的三维结构并进一步认识其生物功能奠定了基础。Objective： To discuss squamous cell carcinomas （SCC） in mucosal system ,including tumors arising from the head and neck, lung, esophagus, and cervix, which accounts for over one third of all cancers and cancer-related deaths annually in the United States. SCC-related oncogene （SCCRO/RP42/DCUN1D1） has been proved exclusively to be amplified in cancer cell lines and involved in Hughog signaling pathway, which is amplified along the 3q26.3 region in human SCC. Methods： In order to know more about the functions of DCUN1D1, firstly the cDNA fragment of DCUN1D1 was amplified from human livers＇ and brains＇ RNA by using RT-PCR. The protein was expressed in soluble form in E.coli and was purified by GST- Sepharose column and Sephadex G-200. Results： The protein DCLrN1D1 was crystallized by the hanging-drop vapour-diffusion method and over 95% purity of protein was obtained as well as the visible crystals under microscope. Conclusions： The growing conditions of the protein crystal DCUN1D1 has been initially found out, which laid a foundation for the analysis of tri-dimensional structure of DCUN1D1 as well as further understanding its biological function.

关 键 词：鳞状细胞癌 DCUN1D1 表达 纯化 晶体生长 

分 类 号：R730.23[医药卫生—肿瘤]