大鼠Occludin基因克隆及其在肺泡Ⅱ型上皮细胞中的表达  

Establishment of rat occludin mammal expression system

作　　者：李玉英[1] 陈枫[2] 谭红梅[3] 王宋平[1] 黄桂君[1] 钱桂生[1] 王关嵩[3] 

机构地区：[1]第三军医大学新桥医院呼吸内科研究所,重庆400037 [2]第三军医大学新桥医院心内科,重庆400037 [3]第三军医大学新桥医院肾病内科,重庆400037

出　　处：《重庆医学》2008年第19期2203-2204,共2页Chongqing medicine

基　　金：重庆市自然科学基金资助项目(2006BB5065)

摘　　要：目的克隆大鼠紧密连接蛋白Occludin基因,构建哺乳动物表达载体。为进一步研究该基因的功能提供手段。方法采用RT-PCR法克隆大鼠Occludin基因编码序列,定向克隆法构建该基因哺乳动物表达载体。脂质体介导该载体转染人肺泡Ⅱ型上皮A549细胞,Western-blot法检测Occludin蛋白表达。结果成功克隆Occludin基因,构建pCMV-Occ哺乳动物表达载体并在A549细胞中表达。结论pCMV-Occ载体可介导Occludin基因在人肺Ⅱ型上皮A549细胞中表达。Objective To clone rat occludin full length cDNA and establish its mammal expression system. Methods RT-PCR method was used in cloning occludin complete code cDNA sequence. Directed cloning was employed in vector construction. Occludin expression in A549 cell line was detected by Western-blot. Results Occludin complete code cDNA sequence was successfully cloned and inserted into the muti clone sites of pCMV-Script. Occludin expression in A549 cell was increased by introducing its mammal expressing vector. Conclusion Occludin mammal expressing vector pCMV-Occ can help increasing expression of occludin in A549 cell line.

关 键 词：OCCLUDIN 克隆 哺乳动物表达载体 

分 类 号：R346[医药卫生—基础医学]