番茄磷酸盐转运蛋白SlPT2基因启动子克隆及功能分析
Cloning and Functional Analysis of Phosphate Transporter SlPT2 Promoter from Solanum lycopersicum
王佳音 1张云逸 1薛婷婷 1闫品月 1朱姝 1李文静2
作者信息
- 1. 廊坊师范学院 生命科学学院,河北 廊坊 065000
- 2. 廊坊师范学院 生命科学学院,河北 廊坊 065000;廊坊市生物样品分析及农残检测重点实验室,河北 廊坊 065000
- 折叠
摘要
为研究番茄磷酸盐转运蛋白SlPT2基因启动子的功能及活性,以番茄Micro-Tom为材料,利用荧光定量PCR检测SlPT2基因表达模式,利用PCR技术克隆SlPT2启动子(p SlPT2)序列,通过启动子在线分析网站Plant Care和New Place分析p SlPT2内部所含有的顺式作用元件,利用同源重组法构建p1300GN-p SlPT2重组载体,转化拟南芥,通过GUS组织化学染色研究SlPT2启动子表达模式及表达活性.荧光定量PCR结果表明,SlPT2主要在番茄根部表达.通过Plant Care和New Place分析发现,启动子(1 801 bp)内部除含有CAAT-Box与TATA-Box等核心启动子元件外,还包含与根特异性表达有关的元件(ROOTMOTIFTAPOX1、RAV1AAT、OSE1ROOTNODULE),以及参与光响应(Box 4、G-Box、TCT-motif)、茉莉酸甲酯反应(CGTCA-motif)、脱落酸反应(ABRE)、玉米醇溶蛋白代谢调控(O2-site)的顺式作用元件等.GUS组织化学染色结果表明,SlPT2启动子驱动GUS基因在拟南芥根部特异表达,将其初步鉴定为根特异性表达启动子.
Abstract
To study the function and activity of the tomato(Solanum lycopersicum)phosphate transporter SlPT2 promoter,tomato variety of Micro-Tom was used as experimental materials,the real-time quantitative PCR was performed to determine the expression pattern of SlPT2 gene,and the sequence of SlPT2 promoter(p SlPT2)was cloned using PCR technology.The cis-acting elements inside p SlPT2 were analyzed through the online promoter analysis websites Plant Care and New Place.The plant expression vector p1300GN-p SlPT2 was constructed using homologous recombination method and transformed into Arabidopsis.The expression patterns and activities of p SlPT2 were studied by using GUS histochemical staining.The real-time quantitative PCR results showed that SlPT2 was expressed mainly in tomato roots;Plant Care and New Place analysis showed that p SlPT2(1 801 bp)not only contained the core promoter elements such as CAAT-Box and TATA-Box,but also contained root-specific expression elements(ROOTMOTIFTAPOX1,RAV1AAT,OSE1ROOTNODULE),light response elements(Box 4,G-Box,TCT-motif),and CGTCA-motif,ABRE,O2-site involved in methyl jasmonate reaction,abscisic acid reaction,regulation of zein metabolism respectively.GUS histochemical staining showed that the SlPT2 promoter drove GUS gene expression in roots of Arabidopsis specifically,which indicated that SlPT2 promoter was a root specific promoter.
关键词
番茄/磷酸盐转运蛋白/组织特异型启动子/GUS组织化学染色/顺式作用元件Key words
Solanum lycopersicum/Phosphate transporter/Tissue specific promoter/GUS histochemical staining/Cis-acting element引用本文复制引用
基金项目
河北省省属高等学校基本科研业务费研究项目(JYQ202101)
出版年
2024
国家科技期刊平台
NSTL
万方数据