百脉根LjIPT3启动子的克隆及表达载体的构建Cloning of LjIPT3 Promoter in Lotus japonicus and Its Expression Vector Construction
杨亚丽;张瑞萍;李美茹;姜华武;吴国江;
摘要(Abstract):
[目的]为深入研究LjIPT3的表达调控和功能奠定基础。[方法]利用TAIL-PCR技术克隆百脉根IPT基因LjIPT3起始密码子的上游序列,利用生物信息学手段对其序列特征和潜在的调控元件进行分析,并构建启动子表达载体。[结果]利用TAIL-PCR技术成功克隆了LjIPT3基因5′端上游调控序列。将目的片段连接到pMD18-T载体上,获长度为1 910 bp序列,其3′端有部分序列与LjIPT3的5′端相同,证明克隆的片段为LjIPT3起始密码子ATG的上游区域。该序列含有1个TATA-box和3个CAAT-box,还包含光响应元件、MYB结合位点、不同的激素反应元件以及各种胁迫元件,说明该基因的表达可能受多种因素的调控。成功构建了驱动报告基因GUS的植物表达载体。[结论]该研究为研究植物细胞分裂素的合成代谢和植物生长发育的调控奠定了基础。
关键词(KeyWords): 百脉根;异戊二烯转移酶;启动子;生物信息学
基金项目(Foundation): 973项目(2001CB1089);; 中国科学院百人计划资助
作者(Author): 杨亚丽;张瑞萍;李美茹;姜华武;吴国江;
Email:
DOI: 10.13989/j.cnki.0517-6611.2008.18.059
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