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拟南芥AtWRKY33基因启动子的克隆及表达分析
童晨晨,张乘,樊婷婷,曹树青
0
(合肥工业大学食品与生物工程学院,合肥 230009)
摘要:
以野生型拟南芥为材料,采用PCR技术克隆得到了拟南芥AtWRKY33基因起始密码子ATG上游1 629 bp启动子序列,并利用该启动子驱动GUS基因在野生型拟南芥中表达,对获得的转基因拟南芥采用重金属Cd处理不同时间,进行GUS染色及定量分析。结果表明:AtWRKY33基因启动子与GUS融合表达载体成功构建并正常启动GUS基因表达;拟南芥植株中的AtWRKY33基因在根中大量表达;定性与定量实验均显示经重金属Cd处理后的拟南芥幼苗中AtWRKY33基因随着时间增加而被显著诱导表达。说明该基因响应重金属Cd胁迫。
关键词:  AtWRKY33基因  启动子  GUS染色  Cd处理  组织特异性表达
DOI:10.13610/j.cnki.1672-352x.20201113.015
投稿时间:2020-02-20
基金项目:国家自然科学基金面上项目(31872803)资助。
Cloning and expression analysis of AtWRKY33 gene promoter from Arabidopsis thaliana
TONG Chenchen,ZHANG Cheng,FAN Tingting,CAO Shuqing
(School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009)
Abstract:
In this study, taken a wild-type Arabidopsis as the experimental material, the promoter fragment in the upstream of the start codon of the AtWRKY33 gene from Arabidopsis thaliana was isolated by PCR cloning method, which was 1 629 bp in length. The promoter was used to drive GUS gene expression in wild Arabidopsis thaliana. The obtained transgenic plants were analyzed by using GUS staining and quantitative assay after different hours of heavy metal Cd stress. The results showed that: the AtWRKY33 gene promoter and GUS fusion expression vector were successfully constructed, and GUS gene expression was normally activated; the AtWRKY33 gene was highly expressed in roots; heavy metal Cd treatment significantly induced the expression of gene AtWRKY33 in a time-rely manner by qualitative and quantitative experiments. In conclusion, the AtWRKY33 gene in Arabidopsis thaliana is response to heavy metal Cd stress.
Key words:  AtWRKY33 gene  promoter  GUS staining  Cd treatment  tissue specific expression

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