一种带有表型标记基因的诱导型抗旱植物表达载体的构建  

Construction of an Inducible Drought-Reisistance Plant Expression Vector with a Phenotype Marker Gene

作　　者：李斯[1] 王爱英[1] 沈海涛[1] 祝建波[1] 

机构地区：[1]石河子大学生命科学学院农业生物技术重点实验室,石河子832003

出　　处：《石河子大学学报（自然科学版）》2009年第6期661-665,共5页Journal of Shihezi University(Natural Science)

基　　金：国家转基因专项(2008ZX08005-004);新疆兵团博士基金(2006jc07)

摘　　要：为了克服组成型表达转录因子基因影响转基因植物性状的缺点,并构建一种具有级联放大作用并带有表型标记的诱导型植物双价表达载体。研究采用PCR方法从拟南芥克隆获得冷诱导转录因子CBF3基因,蜡质合成相关WIN1基因,干旱诱导RD29A基因启动子和冷诱导的LEA14基因启动子,并用CBF3转录因子所调控的下游RD29A基因启动子和LEA14基因启动子分别驱动CBF3基因和WIN1基因表达,构建了双价植物表达载体RD29AP-CBF3/LEA14P-WIN1/pCAMBIA2201。我们预测在转基因植物中,该表达系统可在干旱等逆境信号存在条件下,通过级联放大的方式诱导表达,在增加植物抗逆性的同时,增加叶片表层蜡质的积累,从而易于表型识别。本研究为利用花粉管通道法转化棉花,提高抗逆转基因棉花田间筛选的效率奠定了基础。To overcome the disadvantages of constitutively plant expression vector that makes function by cascade expressing transcription factor genes, an inducible bivalent amplification was constructed. Cold-induced transcription factor CBF3 gene,synthetic wax-related WIN1 gene, the promoter region of drought-induced RD29A and cold-induced LEA14 were cloned from the Arabidopsis by using the PCR method. And we use CBU3 transcription factor which can regulate downstream genes RD29A and LEA14＇s promotion respectively to drive the expression of gene WIN1 and CBF3. We funally constructed the bivalent plant expression vector RD29AP-CBF3/LEA14P-WIN1/pCAMBIA2201. This expression vector might be able to increase the wax accumulation in leaf surface while increasing stress resistance of plants ,depending on the cascade amplification in the transgenic plants which are under drought and other adversity signal conditions, and make the phenotype easily identifiable. This study has laid a foundation of enhancing the screening efficiency to select transgenic cotton which were transformed through pollen-tube pathway in cotton fields.

关 键 词：抗旱 转录因子 诱导型启动子 表型标记 载体构建 

分 类 号：S332.4[农业科学—作物遗传育种]