银杏叶绿素a/b结合蛋白基因(GbLhcb4)及其启动子克隆  

Cloning of light-harvesting cholorophyll a/b-binding protein coding gene(GbLhcb4) and promoter sequence from Ginkgo biloba

作　　者：王欢利[1] 刘新亮[1] 郁万文[1] 李广平[1] 曹福亮[1] 

机构地区：[1]南京林业大学林学院南方现代林业协同创新中心,江苏南京210037

出　　处：《中南林业科技大学学报》2015年第5期114-121,共8页Journal of Central South University of Forestry & Technology

基　　金：国家自然科学基金项目(31170627);江苏省自然基金项目(BK2010019);江苏省普通高校研究生科研创新计划项目(CXZZ12_0546);南京林业大学优秀博士学位论文创新基金项目

摘　　要：为了探究银杏中叶绿素a/b结合蛋白基因(Gb Lhcb4)的结构及其相关的转录调控元件,基于Roche454高通量测序获得的Gb Lhcb4基因片段,采用RACE技术分离得到该基因的c DNA序列,并通过染色体步移技术获得该基因启动子序列,结合生物信息学技术分析基因序列的特征、系统进化地位及启动子区域的转录调控元件。序列分析表明,Gb Lhcb4基因全长1 200 bp,含有一个834 bp开放阅读框,编码277个氨基酸,相对分子量为30.65 k D,等电点为5.17。同源分析表明,该基因编码蛋白与北美云杉Ps Lhcb4蛋白(Gen Bank登录号:ABK21281.1)相似性最高,为74.7%。启动子顺式作用元件预测表明,启动子区域包含多个光响应元件及逆境响应元件,同时还存在激素响应、组织特异性表达、细胞周期等相关调控元件。此研究分离得到银杏Gb Lhcb4基因及启动子序列,并进行了相关的生物信息学分析,为探究银杏Gb Lhcb4基因应答环境信号的转录调控机制提供了分子基础。In order to explore structure and cis-regulatory elements of the gene of light-harvesting cholorophyll a/b-binding proteins in ginkgo （GbLhcb4）, full-length cDNA sequence and promoter of GbLhcb4 were cloned from ginkgo by using method of rapid ampliifcation of cDNA ends and genome walking methods respectively, which based on the fragment of GbLhcb4 acquired by Roche 454 high-throughput sequencing. The sequence analysis indicated that the length of GbLhcb4 （GenBank number：KC818609） was 1200 bp, which contained a 834 bp open reading frame and encoded a 277 aa protein, with a predicted molecular weight of 30.65 kD and the isoelectric point of 5.17. Multiple alignment of deduced amino acid sequence showed that it had the highest sequence similarity, approximately 74.7%similarity with other Lhcb4 in Picea sitchensis. Cis-regulatory elements analysis revealed that light responsiveness elements, stress responsiveness, phytohormone responsiveness and tissue-speciifc cis-regulatory elements was in the promoter region of GbLhcb4. The full-length sequence and promoter of GbLhcb4 were obtained in this research and the structure and cis-elements were analyzed by a series of bioinformatics soft-wares. It will be helpful to understand the transcriptional regulatory mechanism of GbLhcb4, when ginkgo responded to environment signals.

关 键 词：银杏 叶绿素a/b结合蛋白基因 启动子分离 RACE技术 

分 类 号：S792.95[农业科学—林木遗传育种] Q786[农业科学—林学]