摘要
为深入研究巴西橡胶树HbWRKY1基因的表达调控机制,采用染色体步移法克隆了HbWRKY1基因的5′调控区序列。结果表明,该序列具有预测的核心启动子区域位于-6^-57 bp,转录起始位点位于翻译起始位点上游41 bp处;该序列具有TATA-box,CAAT-box和GATA-box等多个典型的真核生物启动子基本的顺式作用元件,同时还具有低温响应元件LTR,赤霉素反应元件GARE-motif,生长素响应元件TGA-element和脱落酸响应元件ABRE等参与激素调控元件,此外还有与MYB结合的MBS元件,热胁迫反应的HSE,厌氧诱导元件ARE和增强诱发厌氧反应的GC-motif等顺式作用元件。构建了不同长度的HbWRKY1启动子片段的植物表达载体并转化本生烟草,通过潮霉素抗性和PCR鉴定,获得了转基因植株。对获得的转HbWRKY1不同长度启动子片段的转基因T1植株GUS染色发现,植株的茎、叶柄、主叶脉和根中均可呈现蓝色,表明HbWRKY1基因的5′调控区可以启动GUS基因的表达,具有启动子的活性。
In order to further study the expression and regulation mechanism of HbWRKY1 in Hevea brasiliensis,the 5′ regulatory region of HbWRKY1 gene from Hevea brasiliensis was cloned using genome walker strategy.Bioinformatics analysis showed that TATA box,CAAT-box and other core configurations were found in this promoter region.Several sequences similar to eukaryotic cis-regulatory elements,such as GARE-motif,TGA element and MBS element,were found in the 5′-UTR proximal 5′flanking sequence of HbWRKY1 gene.Several plant expression vectors with 5′regulatory region of HbWRKY1 gene from Hevea brasiliensis were constructed and transgenic Nicotiana benthamiana plants were obtained by Agrobacterium-mediated transformation.GUS activity analysis revealed that the fragment of 5′ regulatory region of HbWRKY1 drove the expression of the GUS gene transgenic Nicotiana benthamiana plants.
出处
《热带作物学报》
CSCD
北大核心
2013年第4期630-635,共6页
Chinese Journal of Tropical Crops
基金
国家自然科学基金项目(No.31170285)
中央级公益性科研院所基本科研业务费专项资金资助项目(No.ITBB110205)
