摘要
克隆并测序了1.2 kb的鸡cENS-2(chicken normastem 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干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,为下一步对鸡胚胎干细胞(CES)细胞标记研究打下基础。
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.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2005年第10期987-990,共4页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
北京市自然科学基金项目(5051004)
