与内皮抑制素相互作用的蛋白及其抑制血管生成作用的分子机制  

Screening of proteins interacting with endostatin and the molecular mechanism for inhibition of angiogenesis

作　　者：张宝林[1] 范保星[2] 贾向旭[3] 苑晓玲[3] 冯怡[4] 彭善云[3] 马良[1] 刘又宁[2] 

机构地区：[1]解放军总医院科研处,北京市100853 [2]解放军总医院呼吸科,北京市100853 [3]解放军军事医学科学院放射医学研究所,北京市100850 [4]解放军军事医学科学院生物工程研究所,北京市100850

出　　处：《中国临床康复》2005年第38期23-25,i0002,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:探讨与内皮抑制素相互作用的蛋白,阐明其抑制血管生成作用的分子机制。方法:实验于2002-01/2004-12在解放军总医院呼吸科实验室、军事医学科学院放射医学和生物工程研究所实验室完成。应用酵母双杂交系统3,以内皮抑制素为诱饵,筛选人肝cDNA文库,寻找与之相互作用的蛋白。再利用一对一酵母回复性杂交、β-GAL实验和生物信息学分析技术筛除假阳性克隆。结果:成功构建稳定表达内皮抑制素基因的载体;酵母双杂交共获得281个克隆,经验证,6个为阳性克隆,测序发现其中一个克隆为未知序列;FASTCAT试验发现阳性克隆中4个克隆可明显的与内皮抑制素相互作用。结论:6个阳性克隆表达的蛋白可能能与内皮抑制素相互作用,并不同程度的参与了内皮抑制素调节的血管生成过程。AIM： To screen the proteins that interacting with endostatin, and elucidate the molecular mechanism that endostatin inhibits the formation of blood vessel. METHODS： The experiment was carried out in the laboratory of the Respiratory Department, General Hospital of Chinese PLA, and the laboratories of the Radiological Medicine Institute and Biological Engineering Institute of Academy of Military Medical Science, from January 2002 to December 2004. Endostatin was used as bait to screen the human liver eDNA library to find the proteins that interacting with endostatin by using yeast-two-hybrid technique. The false positive clones were screened out by one to one yeast-two-hybrid,β-GAL assay and bioinformatics methods. RESULTS： The vector that can steadily express endostatin gene was successfully constructed, and 281 clones were obtained with yeast-two-hybrid, and furthermore, six clones were verified to be true positive ones. Among the six clones, one was fund to be unknown sequence, and four clones were verified to have an interaction with endostatin by using FAST CAT test. CONCLUSION： The proteins of six positively expressed clones may functionally interact with endostatin and are involved in the process of angiogenesis regulated by endostatin to different extents.

关 键 词：克隆 分子 抑制素 杂交 

分 类 号：R318[医药卫生—生物医学工程]