矮牵牛果实特异表达启动子FBP7的基因克隆及序列分析

为进一步实现外源基因在草莓果实中的特异表达,克隆了矮牵牛果实特异表达启动子FBP7。根据GenBank发表序列设计引物,以矮牵牛(Petunia hybrida)总DNA为模板,通过PCR扩增获得约500 bp的DNA片段,回收该片段并克隆至PUC18载体上,测序后将所得序列与原序列进行比对,其同源性为97%。为分析这些差异序列是否影响启动子功能,进行了启动子功能预测及顺式作用元件分析。结果表明,所克隆的启动子序列应具有启动子功能,且能实现果实特异表达,其序列分析差异可能是由于个体差异及多态性的影响,为草莓果实特异表达启动子的选择提供新思路。

Clone and sequence analysis of fruit specific promoter FBP7 of Petunia hybrida

In order to realize high efficient and specific expression of t-PA in strawberry fruits,FBP7 fruit specific promoter of Petunia hybrida was cloned.The DNA segment of 500 bp was got by PCR amplification using the primer designed according to the sequence in GenBank and the template total DNA extracted from Petunia hybrida.The retrieved segment was cloned to PUC18 vector and sequenced.The sequence was analyzed with DNAMAN software and the homology with primitive sequence was 97%.The function prediction and cis-element analysis were performed in order to analyze whether the sequence disparity would affect the promoter's function.The results indicated that the sequence cloned would function as the promoter and make the specific expression of t-PA gene in fruit.The sequence disparity may come from individual difference and the phenomenon of polymorphism.The results will provide a new choice for promoter specific expression in strawberry fruits.