Isolation and Characterization of CMO Gene Promoter from Halophyte Suaeda liaotungensis K.  

辽宁碱蓬胆碱单加氧酶(CMO)基因启动子分离及功能元件分析(英文)

作　　者：李秋莉[1] 尹辉[1] 李丹[1] 朱宏飞[2] 张毅[1] 朱巍巍[1] 

机构地区：[1]辽宁师范大学生命科学学院,大连116029 [2]辽宁工程技术大学,阜新123000

出　　处：《Journal of Genetics and Genomics》2007年第4期355-361,共7页遗传学报（英文版）

基　　金：This work was supported by the National Natural Sciences Foundation of China (No. 30370806).

摘　　要：The 5＇-flanking proximal region of stress-induced gene encoding choline monooxygenase （CMO） was isolated by Adaptor-PCR and TAIL-PCR from halophyte Suaeda liaotungensis K. A total of 2,204 bp DNA sequence was obtained. The transcription start site, which is located at 128 bp upstream to the start ATG, was predicted by the TSSP-TCM program. The functional elements were analysed by PLACE program. The obtained SICMO gene promoter contains the basic elements： TATA-box, CAAT-box, and stress-induced elements, for example, salt responsive element （GAAAAA）, cold responsive elements （CANNTG）, ABA （Abscisic Acid） responsive elements （NAACAA）, water stress element （CGGTTG）, and WUN responsive elements （GTTAGGTTC）. Isolation and analysis of the promoter of the CMO gene from S. liaotungensis lays a foundation for characterising the stress-induced promoter elements, studying the relationship between the structure and function of the promoter, and investigating the molecular mechanism of CMO gene regulation.根据已知的辽宁碱蓬CMO cDNA 5′端序列设计两个基因特异的反向引物(CR1,CR2),通过衔接头PCR获得了CMO基因起始密码子上游498 bp的序列。根据所获得的序列设计两个基因特异的反向引物(CR3,CR4),用CR2、CR3、CR4分别与4个简并引物配对,通过TAIL-PCR扩增,获得了约2 kb的序列。经Sequencer软件拼接上述两段序列,获得了CMO基因起始密码子上游2,332 bp的序列。用TSSP-TCM软件分析此序列,预测出转录起始点(C)位于起始密码子上游128 bp处,由此我们获得了2,204 bp的SlCMO启动子序列。用PLACE软件分析此序列,发现该序列具有启动子的基本元件TATA-box、CAAT-box,包含多个胁迫诱导元件,如盐诱导元件GAAAAA,冷胁迫诱导元件CANNTG,ABA 响应因子NAACAA,水胁迫元件CGGTTG和伤害诱导元件GTTAGGTTC等,是一个强的胁迫诱导启动子。辽宁碱蓬胆碱单加氧酶基因盐诱导启动子的获得,为盐诱导启动子功能元件分析提供了可能,为进一步研究启动子结构与功能的相互关系、CMO基因的表达调控机制奠定了基础。

关 键 词：Suaeda liaotungensis K. choline monooxygenase （CMO） PROMOTER 

分 类 号：Q943.2[生物学—植物学]