Teng Peng, Cao Ying, Wang Wei-Xian, Zhu Shao-Xia, Meng An-Ming, Zhang Jing-Pu
Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100080, China.
Yi Chuan Xue Bao. 2004 Jan;31(1):39-42.
In vitro experiment showed that HNF3beta 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 HNF3beta 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 HNF3beta binding sites is able to confer the notochord-expressing activity.
体外实验表明，肝细胞核因子3β（HNF3β）是音猬因子（shh）启动子的直接调节因子。为了研究斑马鱼shh启动子在体内的活性，我们构建了表达载体pShh-EGFP，即将一个含有两个HNF3β结合位点的538 bp斑马鱼shh启动子与绿色荧光蛋白（EGFP）连接。将pShh-EGFP DNA显微注射到斑马鱼的单细胞期胚胎中，并用荧光显微镜观察胚胎中绿色荧光蛋白的表达情况。绿色荧光蛋白的表达在原肠胚形成期开始于轴部下胚层。在体节形成期，在脊索中检测到绿色荧光蛋白，但在底板中未检测到。我们的实验表明，含有两个HNF3β结合位点的538 bp shh启动子能够赋予脊索表达活性。
