苦荞查尔酮合酶基因FtCHS1启动子的克隆及分析
发布时间:2018-06-30 11:12
本文选题:苦荞 + 查尔酮合酶基因 ; 参考:《植物科学学报》2017年04期
【摘要】:采用染色体步移技术,从苦荞(Fagopyrum tataricum Gaertn.)中克隆获得Ft CHS1基因5'端侧翼序列,共1038 bp,将其命名为PFt CHS1。生物信息学分析表明,PFt CHS1中A/T碱基含量为63.5%,含有4个可能的转录起始位点,分别位于起始密码子上游-684~-734、-692~-742、-920~-970、-929~-979 bp处,该序列包含TATA-Box和CAAT-Box等启动子核心元件以及与光、低温和激素应答等相关的功能元件。本研究进一步构建了PFt CHS1-p BI101植物表达载体,并瞬时转化拟南芥(Arabidopsis thaliana L.)叶片,结果显示该序列可驱动GUS报告基因的表达。低温(4℃)和光照(UV-B)处理苦荞幼芽后,采用荧光定量PCR技术分析Ft CHS1基因的表达量,结果表明PFt CHS1可响应低温和紫外环境胁迫,从而引起Ft CHS1基因表达量发生变化。
[Abstract]:The chromosome step technique was used to study the genetic properties of Tartary buckwheat (Fagopyrum tataricum Gaertn.) The 5 'flanking sequence of Ft CHS1 gene was obtained, which was named PFT CHS1. Bioinformatics analysis showed that the content of the A / T base in PFT CHS1 was 63.55.It contained four possible transcriptional initiation sites, located at the upper reaches of the initiation codon -684N -734U -692U -742 + -970-920U -970-929N -979 BP. The sequence contained promoter core elements such as TATA-Box and CAAT-Box, as well as light, and light, respectively. Hypothermia and hormone response and other related functional elements. In this study, the plant expression vector PFT CHS1-p BI101 was constructed and transformed into Arabidopsis thaliana (Arabidopsis thaliana L. The results showed that this sequence could drive the expression of Gus reporter gene. The expression of Ft CHS1 gene was analyzed by fluorescence quantitative PCR after treatment with low temperature (4 鈩,
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