摘要： 为了利用PCR技术得到甘蓝型油菜A7-FT基因启动子序列，根据甘蓝型油菜全基因组序列，利用启动子在线预测软件预测其功能与结构，根据其预测的顺式元件的分布，从5'端开始缺失的方式获得5个不同片段长度的启动子序列。构建含不同片段长度启动子的GUS基因表达载体，利用农杆菌介导拟南芥，得到T₂幼苗，经过GUS染色与脱色，探讨A7-FT基因启动子的功能，为研究甘蓝型油菜开花调控机制提供理论基础。通过PCR技术从甘蓝型油菜湘油15号基因DNA中获得A7-FT基因启动子序列。利用PLACE和PlantCARE在线工具对该段序列进行预测，发现A7-FT基因启动子除了存在启动子核心元件CAATbox和TATAbox，还有光应答元件、激素应答元件、胚乳表达应答元件、抗逆性应答元件、生理控制相关的顺式作用元件。基于预测的顺式作用元件的分布情况，设计特异性引物，克隆不同片段长度启动子，与pCAMBIA1303载体构建5'端缺失载体，分别命名为M1、M2、M3、M4、M5。通过农杆菌介导拟南芥，GUS染色与脱色结果显示，在-1 549~-238可能存在一些负调控元件的结合位点，而-238~+1区域是该启动子的核心区段。

关键词: 甘蓝型油菜, FT基因, 表达, A7-FT启动子, 拟南芥

Abstract: Based on the whole genome sequence of Brassica napus, the sequence of A7-FT gene promoter was obtained by PCR. The promoter online prediction software was used to predict the function and structure of the promotor, and five promoter sequences with different fragment lengths were obtained from the 5'-end deletion according to the distribution of predicted cis-elements. The GUS gene expression vector containing promoters with different fragment lengths was constructed and T₂ seedlings were obtained by Agrobacterium -mediated Arabidopsis thaliana. After GUS staining and decolorization, the function of A7-FT gene promoter was explored to study the flowering of B. napus. The study of regulatory mechanisms provided a theoretical basis.The A7-FT gene promoter sequence was obtained from the DNA of Brassica napus L. Xiangyou 15 by PCR. Using the PLACE and PlantCARE online tools to predict the sequence, it was found that in addition to the promoter core elements CAATbox and TATAbox,the A7-FT gene promoter also had photo responses, hormone response components, expression response element of endosperm,response element of stress resistance and physiological control. Based on the predicted distribution of cis-acting elements, specific primers were designed to clone the promoter with different fragment lengths. A 5' segment deletion vector was constructed with pCAMBIA1303 vector and designated as M1, M2, M3, M4, and M5, respectively. Through Agrobacterium -mediated Arabidopsis thaliana, GUS staining and decolorization results showed that there might be some negative regulatory element binding sites between-1 549 and-238, and the core region of the promoter was from-238 to +1.

Key words: Brassica napus L., FT gene, Expression, A7-FT promoter, Arabidopsis thaliana

中图分类号: 

• S635.03

• Q78