白木香查尔酮合成酶基因(AsCHS1)启动子克隆及激素应答元件功能的初步鉴定  

Cloning and Function Analysis of the Chalcone Synthase Gene (AsCHS1) Promoter in Aquilaria sinensis

作　　者：曹天骏 戴好富[2] 李辉亮[2] 郭冬[2] 梅文莉[2] 彭世清[2] 

机构地区：[1]海南大学农学院,海南海口570228 [2]中国热带农业科学院热带生物技术研究所农业部热带作物生物学与遗传资源利用重点实验室,海南海口571101

出　　处：《热带作物学报》2014年第10期1950-1956,共7页Chinese Journal of Tropical Crops

基　　金：公益性行业(农业)科研专项(No.201303117);国家科技支撑计划课题(No.2013BAI11B04);海南省重大科技项目(No.ZDZX2013023-1)

摘　　要：利用染色体步移法克隆了白木香查尔酮合成酶基因（As CHS1）ATG上游1 082 bp的启动子序列,该启动子序列中AT含量高达69.03%,符合真核生物启动子的序列特征。通过启动子预测软件分析可知,该序列的转录起始位点位于翻译起始位点-65 bp处,并且其上游-25-30 bp区域存在TATA-box等典型的真核生物启动子核心元件,同时含有一些顺式作用元件如赤霉素应答元件GARE-motif、茉莉酸甲酯应答元件CGTCA-motif、水杨酸应答元件TCA-element等激素调控元件,光应答元件BoxⅠ、G-Box、ACE,厌氧诱导元件ARE等。通过构建p C-1 082pro As CHS1植物表达载体,借助农杆菌将重组载体转化到烟草叶片中。蛋白定量结果表明,该序列可以驱动GUS的表达,具有启动子活性;脱落酸显著增强该启动子的活性,乙烯则显著抑制该启动子的活性。Based on the Genome Walker strategy,an approximately 1 082 bp sequence was obtained from the DNA of Aquilaria sinensis.Sequence analysis showed that this promoter region contained a TATA-box core element,a CAAT-box,and other cis-acting elements,including an GARE-motif,a TCA element,CGTCA-motif,G-Box,Box I,and a ARE element; these elements are involved in gibberellin responses,salicylic acid responses,methyl jasmonate responses,light response,respectively.The 1 082 bp promoter sequence was inserted in p CAMBIA1381 Z vector to construct a fusion vector containing the promoter connected to the GUS gene.GUS quantitative analysis of transgenic Nicotiana benthamiana carrying the p C-1 082 pro As CHS1 vector showed that the promoter was active in the leaves.It also showed high activity in the group treated by abscisic acid,while restrained activity in the group treated by ethylene.These results suggest that As CHS1 may be regulated by this two phytohormones.

关 键 词：白木香 查尔酮合酶 启动子 染色体步移 瞬时表达 GUS 

分 类 号：Q943.2[生物学—植物学]