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| 鸡cENS-2基因 U3-R序列的克隆分析与表达载体的构建 | |
| 引用本文： | 张传生,耿立英,杜立新.鸡cENS-2基因 U3-R序列的克隆分析与表达载体的构建[J].畜牧兽医学报,2005,36(10):987-990. |
| 作者姓名： | 张传生 耿立英 杜立新 |
| 作者单位： | 1. 中国农业科学院畜牧研究所,北京,100094;河北科技师范学院动物科学系,昌黎,066600 2. 河北科技师范学院生命科学院,昌黎,066600 3. 中国农业科学院畜牧研究所,北京,100094 |
| 基金项目： | 北京市自然科学基金项目（5051004） |
| 摘 要： | 克隆并测序了1．2kb的鸡cENS-2（chicke nnormastem cell gene-2，cENS-2）基因3’端U3-R样结构调控序列。经序列分析和同源性比较表明，所克隆的U3-R序列包含多个转录因子结合位点如AP-1、GATA-1等，同时还包括一个重要的顺式调控元件B区，该区域在鸡正常的胚胎于细胞中有特异的增强子活性。该序列经pMD-18T载体过渡后，定向连接于切去启动子的pEGFP-N1载体的上游，构建了鸡cENS-2基因U3-R样序列调控的绿色荧光蛋白基因报告载体，为下一步对鸡胚胎于细胞（CES）细胞标记研究打下基础。 |
| 关 键 词： | 鸡胚胎于细胞 cENS-2 U3-R 绿色荧光蛋白 |
| 文章编号： | 0366-6964（2005）10-0987-04 |
| 收稿时间： | 2004-07-19 |
| 修稿时间： | 2004-07-19 |
Molecular Cloning and Sequence Analysis of Putative U3-R Sequence of cENS-2 Gene and Construction of Expression Vector |
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| ZHANG Chuan-sheng,GENG Li-ying,DU Li-xin.Molecular Cloning and Sequence Analysis of Putative U3-R Sequence of cENS-2 Gene and Construction of Expression Vector[J].Acta Veterinaria et Zootechnica Sinica,2005,36(10):987-990. | |
| Authors: | ZHANG Chuan-sheng GENG Li-ying DU Li-xin |
| Institution: | 1. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100094, China; 2. Department of Animal Socience, Hebei Normal University of Science and Technology , Changli 066600, China; 3. Department of Life Science, Hebei Normal University of Science and Technology, Changli 066600, China |
| Abstract: | This report describes construction of specific labeling vector in CES cell. The putative U3-R sequence regions of 1.2 kb of chicken Embryonic Normal Stem cell gene 2 was amplified by PCR and successfully sequenced. Based on sequence analysis and alignments of nucleotides sequence, found many factor or protein binding sites such as AP-1, GATA-land an important elements. B region which exhibits a strong enhancer activity in CES and differentiated cells. The CMV promoter was cut off from pEGFP-N1, the putative U3-R fragment was inserted just upstream of GFP. Results of restriction digestion and DNA sequencing showed that the vector for expressing in CES cell was successfully constructed. |
| Keywords: | CES cENS-2 U3-R EGFP |
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