茭白lea3基因全长cDNA的分离克隆及序列分析  

Segregate Cloning and Sequence Analysis of lea3 Gene Complete cDNA in Zizania caduciflora

作　　者：于丽霞[1] 汤晓倩[1] 武晓璐[1] 鄢波[1] 

机构地区：[1]西南林学院园林学院,云南昆明650224

出　　处：《中国蔬菜》2010年第6期14-18,共5页China Vegetables

基　　金：云南省应用基础研究基金项目(2009ZC079M);云南省省部级重点学科;省高校重点实验室及校实验室共享平台资助

摘　　要：第3组胚胎晚期丰富蛋白(LEA3)是一组在胚胎发育后期丰富表达的蛋白,同时它还受水分胁迫和ABA的诱导,与植物抗旱性密切相关。本研究以经过干旱处理的茭白幼苗的mRNA为模板,根据禾本科lea3基因的保守序列设计简并引物进行RT-PCR,得到lea3基因的部分序列,然后结合RACE方法获得全基因两端的序列,经拼接得出茭白lea3基因的全长cDNA,序列分析结果表明该基因的开放阅读框为666bp,编码221个氨基酸,包含2个LEA3基元序列。通过与已报道的LEA3蛋白序列比较,与禾本科中水稻、大麦、小麦、雀麦、玉米的同源性分别为51.2%、41.0%、41.1%、42.0%和40.5%。最终得出结论,茭白lea3基因具有已知物种lea3基因的特征,可以为该基因的功能研究提供参考。目前,该基因已在GenBank中登录,编号为GQ494017。Group 3 later embryogenesis abundant proteins（LEA3）,present in abundance in seeds during the late stages of development,induced by exogenous abscisic acid（ABA）,and associated with drought tolerance.According to high conservative amino acids of reported LEA3 in Gramineae,degenerate primers were designed and a special DNA fragment was obtained from Zizania caduciflora by RT-PCR.Then based on the obtained sequence,some new primers were designed,and two terminals of lea3 gene were cloned by 3＇RACE and 5＇RACE respectively.Splicing the fragments of sequence,the full length cDNA sequence was obtained which indicated that open reading frame region of this gene was 666 bp,coded 221 amino acid,including 2 characteristic motif of lea3 gene only.Through comparing with the reported lea3 genes sequences,we found that Zizania caduciflora shared the identity with Oryza sativa,Hordeum vulgare,Triticum aestivum,Bromus inermis and Zea mays as 51.2 %,41.0 %,41.1 %,42.0 % and 40.5 %,respectively.It was concluded that Zizania caduciflora lea3 gene had genetic characteristics similar with all species and it could provide reference for functional research.At present,this nucleotide sequence data are deposited in GenBank databases under the accession number GQ494017.

关 键 词：茭白 lea3基因 克隆 

分 类 号：S645.2[农业科学—蔬菜学]