梭梭HaFT-3基因启动子的克隆及功能分析

Cloning and functional analysis of HaFT-3 gene promoter from Haloxylon ammodendron

14-3-3蛋白是一种可调节性蛋白,在调控植物生长发育和响应非生物胁迫中发挥了重要作用。使用染色体步移法克隆梭梭14-3-3蛋白基因HaFT-3上游长度1340 bp的启动子序列。PlantCARE和PLACE在线工具预测分析HaFT-3基因启动子,发现大量CAAT-box、TATA-box等基本顺式作用元件及激素响应元件、脱水响应元件等多个与非生物胁迫相关的顺式作用元件。构建融合表达载体pCAMBIA3301-HaFT-3::Intron-GUS,使用浸花法转化拟南芥,并筛选至T3代。不同生长阶段转基因拟南芥GUS组织化学染色显示,HaFT-3基因启动子仅在成苗期的叶片和根部诱导下游基因表达。GUS染色和GUS定量分析显示HaFT-3基因启动子受高温、低温、ABA、IAA诱导。梭梭HaFT-3基因启动子可以响应非生物胁迫,为进一步研究HaFT-3基因功能提供参考。

14-3-3 protein is a kind of regulatory protein,which plays an important role in regulating plant growth and development and responding to abiotic stress.The 1340 bp promoter sequence of Haloxylon ammodendron 14-3-3 protein gene HaFT-3 was cloned by chromosome walking method.According to the prediction analysis of PlantCARE and PLACE,HaFT-3 promoter contains a large number of basic cis-acting elements such as CAAT-box and TATA-box,furthermore,hormone response elements,dehydration response elements and other cis-acting elements related to abiotic stress are also found.The fusion expression vector pCAMBIA3301-HaFT-3::Intron-GUS was constructed and transformed into Arabidopsis thaliana by flower soaking method,and then screened to T3 generation.GUS histochemical staining showed that HaFT-3 promoter only induced downstream gene expression in leaves and roots at seedling stage.GUS staining and GUS quantitative analysis showed that HaFT-3 promoter was induced by high temperature,low temperature,ABA and IAA.HaFT-3 promoter of H.ammodendron could respond to abiotic stress,which provides a reference for further study of HaFT-3 gene function.