草莓FaAN3基因克隆、蛋白纯化及亚细胞定位

Cloning, Protein Purification and Subcellular Localization of FaAN3 Gene from Fragaria × ananassa Duch.

为了探究栽培草莓中FaAN3基因在花青素代谢途径中的功能,本研究以‘红颜’草莓(Fragaria×ananassa‘Benihoppe’)为材料,采用同源克隆的方法分离得到FaAN3基因的完整开放阅读框序列,对该基因进行了生物信息学分析,原核表达和亚细胞定位。结果表明:克隆得到了全长为645 bp的cDNA序列,生物信息学分析发现该序列编码214个氨基酸,预测分子量为22.85 kD,等电点为6.02。FaAN3蛋白为疏水性蛋白,且无跨膜结构域。原核表达后,在上清中获得了纯化后约40 kD的融合蛋白。FaAN3蛋白亚细胞定位信号在细胞核内出现。研究结果可为探明Fa AN3基因在草莓花青素代谢途径中的基因功能提供一定的理论依据。

In order to explore the function of FaA N3 gene in cultivated strawberry in the anthocyanin metabolic pathway, we took Fragaria×ananassa ＇Benihoppe＇ as materials and obtained the open reading frame of FaAN3 gene via homology cloning strategy. Bioinformatics analysis, prokaryotic expression and subcellular location analysis of the gene were carried out. The results indicated that the cDNA sequence containing a 645 bp open reading frame was cloned and obtained. It was found that the sequence encoded 214 amino acids by bioinformatics analysis. The prediction molecular weight and isoelectric point were 22.85 kD and 6.02, respectively. The FaAN3 protein was hydrophobic protein and it did not have transmembrane domain. About 40 kD purified fusion protein was obtained after prokaryotic expression. The subcellular localization signals of FaAN3 protein appeared in cell nucleus. The result of study could provide a theoretical basis for the further research of FaA N3 gene function in the anthocyanin metabolic pathway of strawberry.