兴安落叶松UDP焦磷酸化酶基因启动子的克隆及双元载体的构建  

Cloning of Promoter Sequence of UDP Pyrophosphorylase Gene from Larix gmelinii and Construction of Binary Vector

作　　者：梅丽丽 郭鑫 张尧 林晓飞[2] 张文波[1] Mei Lili;Guo Xin;Zhang Yao;Lin Xiaofei;Zhang Wenbo(Forestry College,Inner Mongolia Agricultural University,Hohhot,010019;College of Life Science,Inner Mongolia University,Hohhot,010021)

机构地区：[1]内蒙古农业大学林学院,呼和浩特010019 [2]内蒙古大学生命科学学院,呼和浩特010021

出　　处：《分子植物育种》2018年第22期7336-7342,共7页Molecular Plant Breeding

基　　金：国家自然科学基金(31660213;31560065;31160143;31260168;31060106);内蒙古自治区科技创新引导奖励基金;内蒙古自然科学基金(2015MS0354);内蒙古农业大学博士基金(BJ08-27)共同资助

摘　　要：为研究兴安落叶松UDP焦磷酸化酶基因(LgUGPase)的表达特性,利用Tail-PCR的技术克隆了1 376 bp的LgUGPase基因上游序列。利用PlantCARE对该序列进行预测与分析发现,其含有TATA-BOX、CAAT-BOX等启动子基本元件,同时包含光元素响应元件,低温和干旱等逆境响应元件,赤霉素、茉莉酸甲酯和生长素等植物激素响应元件。由此推断LgUGPase基因的表达可能受到光、低温、赤霉素、茉莉酸甲酯等的调控。为进一步研究LgUGPase基因的表达模式和调控机理,将该启动子序列克隆到pORE-R1载体GUS基因的上游,构建了pORE-R1-pLgUGPase::GUS双元表达载体,为通过遗传转化拟南芥及GUS染色研究该启动子的功能及LgUGPase的表达特性提供理论参考。To study the expression characteristics ofLgUGPase gene, a 1 376 bp upstream sequence of LgUGPase was cloned using Tail-PCR. Sequence prediction and analysis by the online software PlantCARE showed that the cloned sequence contained TATA-BOX, CAAT-BOX and some regulatory elements, as well as light-responsive elements, stress response elements （low temperature and MYB binding sites involved in drought-inducibility）, and plant hormones response elements （gibberellin, methyl jasmonate, and auxin）. The result suggested that the expression of LgUGPase gene might be regulated by light, low temperature, gibberellin, methyl-jasmonate and the other factors. In order to further study the expression pattern and regulation mechanism ofLgUGPase gene, the promoter sequence was cloned into the upstream of GUS gene of pORE-R1 vector, constructing the binary expression vector of pORE-RI-pLgUGPase：：GUS, which provided the foundation for studying the function of the promoter and expression characteristics of LgUGPase gene by genetic transformation ofA rabidopsis thaliana and GUS staining.

关 键 词：兴安落叶松 UDP焦磷酸化酶基因 启动子 双元表达载体 

分 类 号：S791.222[农业科学—林木遗传育种]