脐橙CsCAB基因的克隆及表达载体构建

Cloning of CsCAB Gene from Navel Orange and Construction of Its Plant Expression Vectors

从脐橙果皮褐变相关基因的cDNA抑制差减文库中,获得了一个CAB基因家族的同源EST序列,通过快速扩增cDNA末端(RACE)方法成功克隆得到984bp的CsCAB全长基因(GenBank登录号EF010854)。序列分析结果表明,CsCAB包含一个621bp的开放阅读框(ORF),编码207个氨基酸;CsCAB蛋白与钙结合蛋白具有很高的同源性,且具有钙结合蛋白的保守结构域EF-hand。并构建了脐橙CsCAB基因的正、反义植物表达载体pCAMBIA1301-CABs和pCAMBIA1301-CABas,为该基因的功能研究奠定了基础。

A suppression subtractive hybridization （SSH）library was constructed to identify differentially ex- pressed genes in peel pitting of navel orange fruit in previous work. Based on the sequence of a cDNA fragment with significant similarity to CAB gene family,the full-length cDNA of CsCAB was isolated by Rapid Amplification of cDNA Ends（RACE）. CsCAB is of a 621 bp open reading frame（ORF） encoding a protein of 207 amino acids. Sequence anal- ysis showed that the CsCAB protein had a strikingly conserved EF-hand. Furthermore,its sense and antisense plant ex- pression vertors, pCAMBIA130-CAB and pCAMBIA130-CABas, were successfully constructed, providing a basis to study the function and regulation of CsCAB gene.