摘要
CEP6 5蛋白 (cancerandembryoexpressionprotein 6 5 )为肿瘤单克隆抗体 3H11所识别的抗原分子 ,RT PCR及RNA印迹检测表明 ,CEP6 5蛋白表达于肿瘤及胚胎组织 .为了研究CEP6 5在肿瘤发生中的作用机制 ,以CEP6 5为诱饵蛋白 ,通过酵母双杂交体系筛选人胚胎组织cDNA表达文库 .从 5 2× 10 6个克隆中筛选得到 14个阳性克隆 ,并在酵母体系中通过不同方法得到验证 .在此基础上 ,选取在双杂交中作用明显的 5 1号克隆 ,与CEP6 5作GSTpull down实验 ,结果证明 ,CEP6 5与 5 1号阳性克隆蛋白确能相互作用 .生物信息学分析发现 ,CEP6 5与 5 1号阳性克隆蛋白相互作用 。
Cancer and embryo expression protein 65 (CEP65) specifically reacted with Mab3H11, which can bind specifically to different cancer cells from different tissues. Northern blot and RT-PCR showed that the mRNA of CEP65 was extensively distributed in embryonic tissue and different cancerous tissues, but not in corresponding normal tissues. In order to study the function of CEP65 in tumor development, using the yeast two-hybrid system, with CEP65 as a bait, a placenta cDNA expression library was screened, and fourteen positive clones was isolated in yeast from 5.2 x 10(6) clones. To ascertain the interaction, the 51st clone which interacted more intensive with CEP65 in yea was selected to do GST pull-down assay with CEP65. The result indicated that CEP65 also interacted with the 51st clone in vitro. Bioinformatics analysis showed the interaction between CEP65 and the 51st clone maybe regulate the cell differentiation and proliferation by WW domain or protein kinase C.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2004年第6期512-517,共6页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金资助项目 (3 0 2 70 685 )
北京市科技计划重大项目 (H0 2 0 2 2 0 0 2 3 10 )
北京大学基因中心项目~~
