摘要
目的:克隆菊花耐盐碱相关锌指蛋白基因,并进行盐胁迫和品种间差异的表达分析。方法:从大量菊花资源中筛选出抗盐碱品系小菊‘阳光’,利用RT-PCR从经150 mmol/L碳酸钠处理的小菊‘阳光’叶片中分离得到一个锌指蛋白cDNA全长克隆,用Northern杂交检测其在不同盐处理和不同品种小菊中的表达。结果:获得了全长794 bp的基因CmSTZF(GenBank接受号为DQ864730),编码区为170个氨基酸残基。Blast分析表明,CmSTZF含有AN1型锌指结构,序列模式为C-X2-C-X(9-12)-C-X(1-2)-C-X4-C-X2-H-X5-H-X-C,由Cys110-Cys113-Cys131-His134及Cys124-Cys126-Cys142-His140分别围绕锌离子与其他氨基酸共同组成2个锌指四面体结构;在第67～76及第94～104氨基酸残基序列间存在核定位信号。同源性比较发现,CmSTZF与水稻OsISAP1具有54%的同源性,而二者的锌指保守区相似性达100%。Cluster分析表明,小菊CmSTZF锌指蛋白与水稻的2种逆境反应蛋白亲缘关系最近,归属同一类逆境功能蛋白。在150 mmol/L碳酸钠胁迫下,耐盐小菊‘阳光’锌指蛋白表达量明显高于非耐盐小菊‘神韵’,表明CmSTZF锌指蛋白基因在盐碱胁迫下起重要的调控作用。结论:克隆了小菊耐盐碱相关的锌指蛋白基因CmSTZF,其在耐盐小菊‘阳光’中的表达量高于非耐盐小菊‘神韵’,这为小菊锌指蛋白基因CmSTZF耐盐碱功能分析奠定了基础。
Objective： To clone stress-tolerance zinc finger gene and detect its expression character in Chrysanthemum morifolium. Methods： The full-lenth cDNA clone was obtained with RT-PCR from ground cover chrysanthemum ＇Sunlight＇ treated under 150 mmol/L Na2CO3 stress, which had been screened in sah-tolerrance from a great number of chrysanthemum species or varies. The expression character of the cloned gene was detected by Northern hybridization. Results： The cDNA named CmSTZF （GenBank accession No.DQ864730） was of 794 bp and contains open reading frames that encode deduced protein with 170 amino acid residues. Blast analysis showed that ANl-type zinc finger region which had a con- sensus sequence of C-X2-C-X（9-12）-C-X（1-2）-C-X4-C-X2-H-X5-H-X-C consisted in CmSTZF sequence. Two tetra- hedron structures of zinc finger protein were formed by Cys^110-Cys^113-Cys^131-His^134 and Cys^124-Cys^126-Cys^142-His^140 surrounded zinc ion. There were two NLS wherein residues from 67 to 76 and from 94 to 104 basic residues. Cluster analysis showed homology to CmSTZF has 54% identity over complete cds and 100% identity over a stretch of zinc finger conserver region to OslSAP1. Cluster analysis showed CmSTZF protein belonged to the same genetic protein famliy with OslSAP1 and putative multiple stress-responsive zinc-finger protein. Under 150 mmol/L Na2CO3 stress, CmSTZF expression of C.morifolium ＇Sunlight＇ （sah-toleranted） was obviously super to one of ＇Venve＇ （free sah-toleranted）. So it was suggested that CmSTZF plays an important role in the action of transcription regulation under salt-tolerant stress. Conclusion： A zinc finger gene CmSTZF was cloned in C.morifolium and its expression was higher in ＇Sunlight＇ （sah-toleranted） than in ＇Venve＇ （free sah-toleranted）.
出处
《生物技术通讯》
CAS
2008年第1期39-42,共4页
Letters in Biotechnology
基金
国家林业局"948"项目(2005-4-35)
国家高技术研究发展计划项目(JY03-B-27)
关键词
小菊
锌指蛋白
克隆
表达
Chrysanthemum morifolium
zinc finger protein
cDNA clone
expression
