白桦BpZFP4基因启动子克隆和逆境响应元件功能分析

Cloning and Functional Analysis of BpZFP4 Promoter from Birch( Betula platyphylla)

前期研究发现白桦锌指蛋白BpZFP4基因响应多种非生物逆境胁迫。为了深入研究其调控机制,本研究利用染色体步移技术克隆了该基因上游1360 bp的启动子区域,序列分析结果表明该区域含有多个逆境响应顺式作用元件、激素响应元件、光响应以及病原菌和损伤响应元件等。在此基础上,构建了BpZFP4启动子5′-端一系列缺失片段融合GUS报告基因的表达载体。通过对系列缺失突变体在正常条件以及渗透胁迫、盐胁迫和脱落酸(ABA)处理条件下的GUS基因表达分析,鉴定得到BpZFP4启动子对干旱、盐和ABA应答响应的主要调节区域及可能的顺式作用元件。本研究结果为进一步探究BpZFP4基因应答非生物胁迫和信号分子刺激的调控机理提供了重要的理论依据。

The BpZFP4 plays significant roles in abiotic stress responses.To better understand the regulatory mechanisms of BpZFP4 gene expression,we cloned a 1360 bp long promoter fragment of BpZFP4 using genome walking technique.Computational analysis affirmed the existence of multiple abiotic stress responsive cis-elements as well as phytohormone,light,pathogen and wound responsive motifs.Deletion analysis of the BpZFP4 gene promoter in transgenic tobacco plants revealed the key regions and the possible cis-element regulating the BpZFP4 response to stresses such as drought,salt and ABA stress.This result may provide an important contribution for elucidating the regulation mechanisms of BpZFP4 expression in responding to abiotic stress and signalling molecule.