斑马鱼shh启动子指导绿色荧光蛋白基因在脊索中的表达  

The shh Promoter of Zebrafish Directs the Expression of GFP in Notochord

作　　者：滕鹏[1] 曹莹[2] 王为先[1] 朱少侠[1] 孟安明[2] 张靖溥[1] 

机构地区：[1]中国科学院遗传与发育生物学研究所中,北京100080 [2]清华大学生物科学与技术系,北京100084

出　　处：《Acta Genetica Sinica》2004年第1期39-42,共4页

基　　金：国家重点基础研究发展规划 ( 973 )项目 (编号 :G2 0 0 0 0 1610 8);国家自然科学基金 (编号 :3 0 0 2 5 0 2 0 )资助项目~~

摘　　要：体外实验业已证明shh基因的启动子受HNF3β蛋白直接调控。为研究斑马鱼shh启动子在体内的作用模式 ,构建了由 5 38bp斑马鱼shh启动子 (包含两个HNF3β结合位点 )与绿色荧光蛋白EGFP组成的表达载体 ,命名为pShh EGFP。将pShh EGFP注射到斑马鱼 1 细胞期内的受精卵中 ,定期在倒置荧光显微镜下观察GFP的表达。GFP在原肠作用期就开始表达 ,主要发生在中轴下胚层中 ;在体节形成期 ,GFP在脊索细胞中表达 ,但未发现在神经底板细胞中表达。由此可见 ,含两个HNF3β结合区域的 5In vitro experiment showed that HNF3β was the direct regulator of sonic hedgehog (shh) promoter.To investigate the activity of zebrafish shh promoter in vivo,we constructed the expression vector pShh-EGFP with ligating a 538 bp zebrafish shh promoter,which contained two HNF3β binding sites,to EGFP.The pShh-EGFP DNA was microinjected into one-cell stage embryos of the zebrafish and the embryos were observed for GFP expression with fluorescent microscopy.GFP expression started during gastrulation in the axial hypoblast layer.During segmentation,GFP was detected in the notochord but not in the foor plate.Our experiment demonstrated that the 538 bp shh promoter containing two HNF3β binding sites is able to confer the notochord-expressing activity.

关 键 词：SHH 启动子 HNF3β 脊索 斑马鱼 胚胎 绿色荧光蛋白 

分 类 号：Q953[生物学—动物学]