摘要
海岛棉GbMYB5基因属于MYB家族转录因子,该家族基因可能参与调控与生物或非生物逆境相关的一系列基因表达。为了进一步研究该基因的功能,利用PCR技术从棉花基因组DNA中分离到GbMYB5基因上游2.2 kb的启动子序列。软件分析和预测结果表明,该序列具有典型的启动子结构,并含有多种光响应元件、脱落酸和赤霉素等激素响应顺式作用元件、干旱等逆境胁迫响应顺式作用元件以及水杨酸响应顺式作用元件。将克隆到的启动子序列置换PBI101中的CaMV35S启动子,驱动其下游的GUS基因,构建植物表达载体pGbMYB5-GUS,通过花序浸染法转化拟南芥,获得转基因植株。组织化学染色结果表明,在GbMYB5启动子的驱动下,报告基因GUS在拟南芥的根部、叶尖部及生长点均有表达,因此该启动子可能是一个组织特异性启动子。
GbMYB5 gene of Gossypium barbadense L.belongs to the big transcript factor family of MYB genes,which may participate in a series of gene expression related to biotic and abiotic stresses.To further study the function of GbMYB5 gene,a 2.2 kb promoter sequence from G.barbadense L.was isolated by PCR,which belongs to upstream DNA fragment of GbMYB5 gene.Sequence analysis revealed that the sequence was the promoter of GbMYB5 gene,and contained a variety of cis-elements responding to light signaling pathways（Box-I,Box-4,I-box,G-Box）,phytohormones（ABA,GA,ethylene）,stress（heat,drought） and cis-acting element involved in salicylic acid responsiveness（TCA-element）.The plant expression vector with GUS reporter gene（pGbMYB5-GUS） driven by GbMYB5 promoter was constructed from PBI101 by substituting the CaMV35S promoter,and the vector containing GbMYB5 promoter was transferred into Arabidopsis thaliana using Floral-dip method.Histochemical staining revealed that GUS gene was exclusively expressed in root,blade tip,and apical meristem in transgenic Arabidopsis thaliana,which indicated GbMYB5 promoter was a tissue-specific promoter.
出处
《江苏农业学报》
CSCD
北大核心
2012年第5期957-962,共6页
Jiangsu Journal of Agricultural Sciences
基金
国家重大专项(2009zx08005-003B)
江苏省农业科技自主创新基金项目[CX(11)1020]
国家自然科学基金项目(31101746/C170108)
关键词
海岛棉
GbMYB5基因
启动子
拟南芥
表达分析
Gossypium barbadense L.
GbMYB5 gene
promoter
Arabidopsis thaliana
expression analysis
