摘要： 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。
关键词:
脐橙,
CsCAB,
克隆,
表达载体,
构建
Abstract: A suppression subtractive hybridization(SSH)library was constructed to identify differentially expressed 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 analysis showed that the CsCAB protein had a strikingly conserved EF-hand.Furthermore,its sense and antisense plant expression vertors,pCAMBIA130-CAB and pCAMBIA130-CABas,were successfully constructed,providing a basis to study the function and regulation of CsCAB gene.
Key words:
Navel orange,
CsCAB,
Cloning,
Exp ression vector,
Construction
中图分类号:
苏丽艳, 李正国, 樊晶, 高雪, 杨迎伍, 邓伟, 郝彦伟. 脐橙CsCAB基因的克隆及表达载体构建[J]. 华北农学报, 2010, 25(2): 40-43. doi: 10.7668/hbnxb.2010.02.008.
SU Li-yan, LI Zheng-guo, FAN Jing, GAO Xue, YANG Ying-wu, DENG Wei, HAO Yan-wei. Cloning of CsCAB Gene from Navel Orange and Construction of Its Plant Expression Vectors[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2010, 25(2): 40-43. doi: 10.7668/hbnxb.2010.02.008.