IDO-EGFP表达载体的构建及其在人软骨细胞中的表达  

Construction of mammalian expression vector for IDO and EGFP fusion protein and its expression in human chondrocytes

作　　者：何贤辉[1] 徐丽慧[1] 刘毅[1] 江逊[1] 蔡小嫦[2] 曾耀英[1] 

机构地区：[1]暨南大学组织移植与免疫教育部重点实验室,广东广州510632 [2]暨南大学第一附属医院皮肤科,广东广州510632

出　　处：《免疫学杂志》2004年第3期172-176,共5页Immunological Journal

基　　金：国家重点基础研究发展规划项目 (G1 9990 5430 3;G2 0 0 0 570 0 6);国家自然科学基金重点项目 (30 2 30 350 );广东省"十五"科技计划重大专项基金 (A30 2 0 2 0 2 0 4 )资助

摘　　要：目的　克隆人吲哚胺双加氧酶 (IDO)基因并构建IDO与增强型绿色荧光蛋白 (EGFP)融合蛋白哺乳动物细胞表达载体。方法　利用RT PCR方法从人白细胞克隆IDO基因并构建IDO EGFP融合蛋白表达载体。通过细胞核转染仪将表达载体转入人软骨细胞进行瞬时表达 ,以共聚焦显微镜对表达的绿色荧光进行分析。结果　从活化的人白细胞中克隆到IDO基因编码区全长 ,并构建了IDO EGFP融合蛋白表达载体。以该表达载体转染原代人软骨细胞 ,表达的融合蛋白较均匀地分布于整个细胞。结论　成功构建了IDO EGFP哺乳动物细胞表达载体 。Objective To construct a mammalian cell expression vector for human indoleamine 2,3-dioxygenase (IDO) fused with enhanced green fluorescent protein (EGFP). Methods Human IDO gene was cloned by RT-PCR from human leukocytes and was then inserted into pEGFP-N1 vector to construct expression vector for IDO-EGFP fusion protein. The expression of the fusion protein in human chondrocytes transfected with this vector was analyzed by confocal microscopy. Results The full-length coding sequence of IDO was cloned from activated human leukocytes and the expression vector for IDO-EGFP fusion protein was constructed. The expression vector was transfected into human primary chondrocytes and confocal microscopy observation showed that the expressed fusion protein was distributed evenly in the cells. Conclu- sion The expression vector for IDO-EGFP fusion protein has been constructed successfully. These results pave the way for further study of IDO.

关 键 词：吲哚胺双加氧酶 绿色荧光蛋白 融合蛋白 软骨细胞 共聚焦显微术 

分 类 号：R392[医药卫生—免疫学]