《植物生理学报》 2010, 46(2): 113-119

西伯利亚蓼钙调蛋白基因的克隆及其在盐胁迫下的表达

刘春, 刘关君*, 曲春浦, 魏志刚, 刘桂丰, 杨传平

东北林业大学林木遗传育种与生物技术教育部重点实验室, 哈尔滨150040

通信作者：刘关君；E-mail: liuguanjun2003@126.com；Tel: 0451-82190 607

摘　要：

已从西伯利亚蓼叶中 cDNA 文库中获得的钙调蛋白 EST 序列, 采用 cDNA 末端快速扩增(RACE)技术克隆了具有完 整编码区的钙调蛋白基因的 cDNA 序列(GenBank 登录号 GQ988382), 命名为 PsCaM。该基因全长 615 bp, 编码区为 450 bp, 编码 149 个氨基酸, 5' 非翻译区为 63 bp, 3' 非翻译区为 102 bp。同源性分析表明, 该蛋白与其他植物钙调蛋白高度保守, 氨基酸同源性高达 98%。用实时荧光定量 PCR 研究 3% NaHCO3 胁迫下西伯利亚蓼基因表达的结果显示, 自然条件下, 该 基因在叶中表达量最高, 地下茎次之, 茎中最低; 盐胁迫下 CaM 在西伯利亚蓼的地下茎、茎和叶中均有表达, 表达模式不 同。

关键词：钙调蛋白; 西伯利亚蓼; 基因克隆; 实时荧光定量PCR; 表达分析

收稿：2009-10-15   修定：2009-12-28

资助：教育部科学技术研究重点项目(109054)。

Cloning of CaM Gene from Polygonum sibiricum Laxm. and Its Expression under Salt Stress

LIU Chun, LIU Guan-Jun*, QU Chun-Pu, WEI Zhi-Gang, LIU Gui-Feng, YANG Chuan-Ping

Laboratory of Forest Genetics and Breeding and Biotechnology of Ministry of Education, Northeast Forestry University, Harbin 150040, China

Corresponding author: LIU Guan-Jun; E-mail: liuguanjun2003@126.com; Tel: 0451-82190 607

Abstract:

PsCaM gene (GeneBank accession No. GQ988382) containing a complete ORF was obtained using rapid amplification of cDNA ends (RACE) method based on EST sequence which was obtained from cDNA library of Polygonum sibiricum leaves. The full length of cDNA was 615 bp with a 450 bp open reading frame (ORF) which encoded a peptide of 149 amino acids and containing a 63 bp 5' UTR and a 102 bp 3' UTR. The results of homologous sequence analysis indicated that PsCaM was highly conserved in plants, which shared 98% homology with other CaM in the amino acid sequence. Expression analysis of RT-PCR showed that PsCaM was expressed in leaves, stems and rhizomes. The expression level of PsCaM gene was higher in leaves bthan that in rhizomes, and lowest in stems under nature condition. The expression pattern of PsCaM gene was different in leaves, stems and rhizomes under NaHCO₃ stress.

Key words: CaM; Polygonum sibiricum; gene cloning; RT-PCR; expression analysis