摘要： 为了进一步提高转BADH基因植株的胁迫抗性,分离强胁迫诱导型启动子,以生长在新疆北部干旱沙漠地带的异苞滨藜(Atriplex micrantha)为研究对象,依据已经克隆的BADH基因cDNA序列,参照陈柏君等的方法,分离到长为1 509 bp的5’端上游序列(Genebank:Hm30828),预测其转录起始位点为T(+1),位于ATG翻译起始位点的上游82 bp处,同时还发现了一段新的内含子序列(260 bp)。经PLACE和PLANT CARE软件分析,发现此序列具有启动子的基本转录元件:TATA盒和CAAT盒,包含多个抗逆调控元件:ABA响应元件(ABRE),水杨酸诱导元件(-58bp),热激(HSE)、低温(LTRE1)、干旱(DRE)、伤害诱导元件(WUN)等。并首次发现许多其他的胁迫诱导元件:铜胁迫元件(Cu)、真菌诱导元件(AM)、糖信号转导元件(Sugar)、厌氧和低二氧化碳调控元件,植物激素响应元件(ARR1)以及多个光调控元件(Light)和转录因子响应元件(WRKY、CBF、MYB),因此,推测分离到一个新的强胁迫诱导型启动子。同时构建了BADH-P-a-3301和BADH-P-b-3301两个植物表达载体。

关键词: BADH基因, 启动子, 内含子, 克隆

Abstract: In order to further improve the tolerance of BADH transgenic plants and obtain stronger stress-induced promotor,a regulatory sequence of 1 509 bp(Genebank:HM230828) was isolated from Atriplex micrantha,which grows in arid desert Xinjiang,according to the BADH cDNA sequences and refer to the methods of Chen.The transcription start site,which localized at 82 bases upstream of the start ATG,was predicted by soft-berry,a new intron(260 bp)was found.The functional elements were analysed by PLACE and PLANT CARE programm.The promotor contains lots of stress-induced elements,such as ABA response elements,salicylic acid,heat stock,low temperature,drought and wounding response elements,etc.Cu-stress,AM,Sugar,Anaerobic regulatory elements,GA response element,a number of light regulatory and transcription factor response elements(WRKY,CBF,MYB) was discovered firstly.We speculate that it is a new strong stress-induced promoter,we also constructed two plant expression vectors,BADH-P-a-3301 and BADH-P-b-3301.

Key words: BADH gene, Promotor, Intron Clone

中图分类号: 

• Q78