盐穗木盐相关转录因子HcSCL13基因启动子的克隆及活性初步分析

doi:10.13560/j.cnki.biotech.bull.1985.2017.05.019

摘要/Abstract

摘要： 为探明盐穗木盐相关转录因子基因HcSCL13的表达调控规律,利用基因组步移法成功克隆获得该基因2 200 bp 的启动子序列。PlantCARE数据库分析结果表明,该启动子不仅含有启动子区的核心元件 CAAT-box和 TATA-box,还包含多个与逆境应答有关的顺式调控元件。将克隆获得的HcSCL13转录因子基因启动子序列定向替换pBI121载体上的35S启动子,构建融合表达载体并转染模式植物拟南芥,对转基因拟南芥进行GUS组织化学染色。结果显示转基因拟南芥整株被染色,提示该启动子具有表达活性且可能为组成型启动子。

关键词: 盐穗木转录因子HcSCL13基因, 染色体步移, 启动子克隆, 元件及启动活性分析

Abstract: In order to explore the expression characteristic and function of GRAS transcription factor gene HcSCL13 from Halostachys caspica,the 2 200 bp sequence of promoter was cloned using the genomic walking method. The analysis results using the PlantCARE software suggested that the promoter sequence contained not only the core elements such as CAAT-box and TATA-box,but also some cis-element related to the stress response. The 35S promoter sequence of the pBI121 expression vector was replaced by the HcSCL13promoter sequence to construct the fusion expression vector. Then,the fusion expression vector was transformed into Arabidopsis thaliana by flora method and the T1 seedlings were stained by GUS. The results of GUS staining showed that the whole transformed A. thalianaseedlings were stained,indicating that the HcSCL13 gene promoter had expression activity,and it might be constitutive promoter.

Key words: transcription factor gene HcSCL13, genomic walking method, promoter cloning, analysis of cis-acting element and promoter activity

樊寿德,王艳. 盐穗木盐相关转录因子HcSCL13基因启动子的克隆及活性初步分析[J]. 生物技术通报, 2017, 33(5): 131-138.

FAN Shou-de WANG Yan. Cloning and Activity Analysis of Promoter of GRAS Transcription Factor Gene HcSCL13 from Halostachys caspica[J]. Biotechnology Bulletin, 2017, 33(5): 131-138.

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