草莓CO基因克隆与表达分析

Cloning and expression analysis of CONSTANS gene in strawberry

采用RT-PCR和RACE技术从短日照草莓品种‘卡姆罗莎’(Fragaria×ananassaDuch.cv.‘Cama-rosa’)嫩叶中克隆得到与草莓成花相关的基因,命名为FaCO(GenBank登录号,FJ377616)。该基因cDNA全长1475 bp,1 152 bp的开放阅读框编码一个含有384个氨基酸,分子质量为42 ku的蛋白。该蛋白具有典型的CO同源蛋白结构,包含B-box1,B-box2,CCT结构域及COOH区域。序列分析结果表明,草莓CO蛋白与其他物种的CO蛋白具有较高的同源性,其中与杨树(Populus deltoids)的CO蛋白同源性最高,差异为80%。构建pGEX-4T-1/FaCO原核表达体系,获得了分子质量为60 ku的融合蛋白。

A CONSTANS homologous gene named FaCO was cloned from young leaves of strawberry（Fragaria ×ananassa Duch.cv.＇Camarosa＇） by RT-PCR and RACE technology.The gene had a cDNA sequence with 1 475 bp in length and contained an open reading frame of 1 152 bp which coding a deduced 384 amino acids protein.The peptide had two B-boxes,a CCT domain and a COOH region shared highly homology with CO of other plants,especially with CO of Populus deltoids,to 80%.The pGEX-4T-1/FaCO prokaryotic expression systems were constructed and induced.SDS-PAGE analysis showed that the rFaCO molecular weight was about 60 ku.