畜牧兽医学报2012，Vol.43Issue(4)：546-552,7.
鸭CD8α基因启动子区的克隆及荧光素酶报告基因重组体的构建
Molecular Cloning and Construction of Luciferase Reporter Recombinant of CD8α Gene Promoter in Duck
摘要
Abstract
This experiment was conducted to explore the potential regulatory sequences of duck CD8α gene and the mechanism of transcription regulation. The CD8α gene promoter was cloned by genome walking, the sequence of the promoter was analyzed by online software of bioinforma-tics, and then the promoters from DHV-resistant and DHV-susceptible duck were subcloned into the luciferase expression vector pGL3-Basic directly, and their luciferase activity was detected. The DNA sequence of 2 480 bp was amplified including 56 bp in exonl and 2 424 bp of promoter region. There were TATA-box, GC-box, CAAT-box and transcriptional start site in the sequence. 41 transcriptional factor binding sites including CdxA, Nkx-2, GATA-1, SRY, and so on, were detected. The luciferase reporter gene recombinant pGL3- CD8<z including correct target gene was constructed and identified by restrictive endonuclease enzyme cutting and sequencing,the pGL3-Gy and pGL3-Gk plamids had strong luciferase activity. The result lay a foundation for exploring the promoter activity and transcriptional regulation mechanism of duck CD8α gene.关键词
鸭/CD8α基因/启动子/荧光素酶报告基因Key words
duck/ CD8α gene/ promoter/ luciferase reporter gene分类
农业科技引用本文复制引用
徐琪,陈国宏,陈阳,黄正洋,赵文明,张扬,李欣钰,赵荣雪,李秀,段修军..鸭CD8α基因启动子区的克隆及荧光素酶报告基因重组体的构建[J].畜牧兽医学报,2012,43(4):546-552,7.基金项目
国家自然科学基金(31101704) （31101704）
现代农业产业技术体系建设专项资金(nycytx-45-04) （nycytx-45-04）
江苏省属高校自然科学基础研究面上项目(07KJB230138) （07KJB230138）
