不结球白菜氯离子通道蛋白ClC-d基因的克隆及表达分析  

Cloning and expression pattern of chloride ion channel protein gene ClC-d in Brassica chinensis L.

作　　者：赵瑞丽[1] 钟凤林[1] 林义章[1] 高世超[1] 叶丽萍[1] 林琳琳[1] 

机构地区：[1]福建农林大学园艺学院,福建福州350002

出　　处：《河北农业大学学报》2014年第2期23-30,共8页Journal of Hebei Agricultural University

基　　金：福建省大宗蔬菜产业体系;福建省自然科学基金"铜胁迫对小白菜的基因差异研究"(2012j01082);福建农林大学校重点建设项目(6112c0409)

摘　　要：根据从不结球白菜中获得的与铜胁迫相关的氯离子通道蛋白基因ClC-d的TDF片段设计引物,运用RACE技术克隆出具有完整阅读框的不结球白菜氯离子通道蛋白基因ClC-d,该基因全长2 752bp,开放阅读框为2 376bp,编码792个氨基酸,分子量为87.06kD,理论等电点为8.35,为碱性蛋白。结构分析表明:该蛋白含有1个电压门控的氯离子通道和2个CBS结构域。进化树分析显示,不结球白菜氯离子通道蛋白基因ClC-d推导的氨基酸序列与拟南芥氯离子通道蛋白基因的相似度高达100%,属于ClC类氯离子通道蛋白。RT-PCR结果表明在铜胁迫下该基因表达量增加,处理10d时达到最高,推测其可能参与对铜胁迫的应答反应。本研究结果有助于进一步研究该基因在不结球白菜逆境应答中的作用及其作用机制。The primers were designed according to the transcript derived fragment from cDNA- AFLP technique which associated with chloride ion channel protein gene ClC-d. The 2 752 bp full length cDNA of chloride ion channel protein gene ClC-d was isolated by rapid amplification of cDNA ends （RACE）, including a 2 376 bp open reading frame encoding 792 amino acid resi- dues. The molecular weight of deduced protein was 87.06 kDa with a theoretical pI of 8.35. The protein has a voltage-gated chlorine ion channel and two cystathionine-beta-synthase, and belong to C1C subfamily. Phylogenetic analysis showed that chloride ion channel protein gene ClC-d had 100% similarity with Arabidopsis thaliana. Real-time PCR revealed that the ex- pression of chloride ion channel protein gene ClC-d was increased and reached the highest until 10 days under copper stress, suggesting that it might response to copper stresses. The resultswould be useful to the study of its functions and mechanism in copper stress responses of Bras- sica chinensis L.

关 键 词：不结球白菜 氯离子通道蛋白 RACE RT-PCR 

分 类 号：S634.3[农业科学—蔬菜学]