白桦W-box元件的载体构建及驱动GUS基因表达分析

The Vector Construction of W-box Cis-acting Element and Analysis of Its Driving GUS gene Expression of Betula platyphylla

植物体内的启动子含有多种顺式作用元件,通过调控下游基因的表达,影响植物的生长发育。本研究以白桦中含有BplMYB46启动子的质粒为模板,将含有W-box元件的启动子短片段克隆出来,经过酶切、连接和热激转化到大肠杆菌中。经过PCR检测及测序分析,结果表明pCAMBIA1301-W-box-GUS重组载体构建成功。获得工程菌后,瞬时侵染烟草后进行GUS染色,结果显示,烟草叶片能被染色,但颜色较浅,说明W-box元件能够驱动GUS基因表达,但驱动能力较低。本研究为后续分析上游转录因子与W-box顺式作用元件的互作以及筛选诱导型启动子从而改良白桦品质提供前期基础数据。

Many cis-acting elements present on plant promoters can affect the plant growth and development by regulating the expressions of downstream genes.In this study,the promoter short fragment containing the W-box element was cloned,the plasmid containing the BplMYB46 promoter of birch as template of PCR,which was transformed into Escherichia coli,after enzyme digestion,ligation and heat shock.The results showed that recombinant vector of pCAMBIA1301-W-box-GUS was constructed successfully,through PCR detection and sequencing analysis.The engineered bacteria was obtained,and transiently transformed into tobacco,then the tobacco leaves GUS staining was performed.The results indicated that tobacco leaves can be stained,but the color is light,which demonstrated that the W-box element can drive the GUS gene expression,but the driving ability is low.The present study will provide basic data for the further analysis of the interaction between upstream transcription fac tors and cis-acting element ofW-box and screening of inducible promoters to improve the varieties of Betula platyphylla.