小麦“陕253”低分子量谷蛋白基因的克隆与序列分析  

Cloning and Sequence Analysis of LMW-GS Genes from Wheat Variety "Shaan 253"

作　　者：李艳亮[1] 高翔[1,2,3] 陈其皎[1,2,3] 董剑[1,2,3] 赵万春[1,2,3] 王明霞[1] 李敏[1] 陈瑞佶[1] 庞红喜[1,2,3] 李哲清[1,2,3] 刘俊[1,2,3] 

机构地区：[1]西北农林科技大学农学院,陕西杨凌712100 [2]陕西省小麦工程技术研究中心 [3]陕西省小麦新品种培育工程研究中心,陕西杨凌712100

出　　处：《麦类作物学报》2010年第2期203-209,共7页Journal of Triticeae Crops

基　　金：陕西省“13115”科技创新工程重大项目(2007ZDKG-01);现代农业产业技术体系建设专项(NYCYTX-001)

摘　　要：为了挖掘和有效利用陕253中的优良基因,根据LMW-GS编码基因5′和3′端的保守序列设计引物,采用PCR方法以陕253的基因组DNA为模板进行扩增,分别回收并克隆到载体上,测序后得到11个新LMW-GS基因。GenBank登录号分别为GQ892576~GQ892580和GQ892583~GQ892588,其长度为903~1 081 bp。其中,GQ892576、GQ892585、GQ892586、GQ892587和GQ892588具有完整编码区,可分别编码305、351、298、351和307个氨基酸残基的成熟蛋白。而GQ892577、GQ892578、GQ892579、GQ892580、GQ892583和GQ892584由于在编码区内出现提前终止密码子,推测为假基因。推导的氨基酸序列结果显示：它们都具有LMW-m型低分子量谷蛋白亚基的典型结构特征。In order to find and utilize the advantageous genes of Shaan 253 and provide more important information for the further genetic improvement.The full-length coding region（open reading frame,ORF） of low-molecular-weight glutenin subunit（LMW-GS） gene was amplified from cultivar ＂Shaan253＂ by using the primer pairs designed according to the conservative sequences of 5＇and 3＇terminal domains.The amplified DNA fragment was separated and recovered from agarose gel,subsequently cloned into pMD19-T vector,and then transformed into E.coli strain DH5α.eleven positive clones（GenBank accession number.GQ892576-GQ892580,GQ892583-GQ892588） were selected and sequenced.these genes possessed the nucleotide sequences from 903bp to 1 081bp.GQ892576,GQ892585,GQ892586,GQ892587 and GQ892588 had the whole CDS,and encoded a mature protein of 305、351、298、351 and 307 amino acid residues,respectively.GQ892577,GQ892578,GQ892579,GQ892580,GQ892583,GQ892584 were putative pseudogenes due to the presence of inframe stop codons in these coding region.Deduced amino acid sequence showed these sequences were the typical structure features of LMW-m type.

关 键 词：小麦 陕253 低分子量谷蛋白 基因克隆 序列分析 

分 类 号：S512.1[农业科学—作物学] S331