花椰菜ERF114基因的克隆及Gateway技术构建表达载体

Cloning of Cauliflower ERF114 Gene and Construction of Its Expression Vector by Gateway

【目的】为进一步探究花椰菜ERF114基因功能,从花椰菜中克隆ERF114基因并构建植物表达载体。【方法】利用RTPCR技术从花椰菜中分离ERF114基因;通过生物信息学软件对ERF114蛋白特性进行预测和分析;采用Gateway技术构建ERF114基因的植物表达载体。【结果】克隆获得了花椰菜ERF114基因,其全长为681 bp,编码226个氨基酸,具有1个AP2结构域。生物信息学分析显示,该蛋白质为亲水性蛋白,有1个跨膜区域,二级结构以α-螺旋,β-折叠和无规则卷曲结构为主。进化树分析表明,花椰菜ERF114与油菜的亲缘关系最近。同时成功构建了植物表达载体。【结论】成功克隆了花椰菜ERF114基因,对其进行生物信息学分析,并构建其表达载体,为探究花椰菜ERF114基因功能及分子调控机制奠定了基础。

[ Objective ] In order to further explore the function of cauliflower ERF114 gene, ERF114 gene was cloned from cauliflower, and its plant expression vector was constructed. [ Method] ERFll4 gene from cauliflower by RT-PCR technology was isolated, and the character- istics of ERF114 protein by bioinformatics software were predicted and analyzed, and the plant expression vector of ERF114 gene by Gateway technology was constructed. [ Result] ERFll4 gene of cauliflower was cloned, and its length was 681 bp, encoding 226 amino acids with a AP2 domain. Bioinformatics analysis showed that the protein encoded by the gene was a hydrophilic protein with a transmembrane domain, and the secondary structure was mainly composed of alpha helix, beta fold and irregular coiled structure. Phylogenetic tree analysis showed that the relationship between cauliflower ERF114 and rape was the closest, meanwhile its plant expression vector was successfully construc- ted. [ Conclusion]The cauliflower ERFll4 gene was successfully cloned, its bioinformatics was analyzed, and its expression vectors were constructed, which laid a solid foundation for further exploring the function and molecular mechanism of cauliflower ERFll4 gene.