麦类作物学报
麥類作物學報
맥류작물학보
JOURNAL OF TRITICEAE CROPS
2010年
2期
203-209
,共7页
李艳亮%高翔%陈其皎%董剑%赵万春%王明霞%李敏%陈瑞佶%庞红喜%李哲清%刘俊
李豔亮%高翔%陳其皎%董劍%趙萬春%王明霞%李敏%陳瑞佶%龐紅喜%李哲清%劉俊
리염량%고상%진기교%동검%조만춘%왕명하%리민%진서길%방홍희%리철청%류준
小麦%陕253%低分子量谷蛋白%基因克隆%序列分析
小麥%陝253%低分子量穀蛋白%基因剋隆%序列分析
소맥%협253%저분자량곡단백%기인극륭%서렬분석
Wheat%Shaan 253%LMW-GS%Gene cloning%Sequence analysis
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.
為瞭挖掘和有效利用陝253中的優良基因,根據LMW-GS編碼基因5'和3'耑的保守序列設計引物,採用PCR方法以陝253的基因組DNA為模闆進行擴增,分彆迴收併剋隆到載體上,測序後得到11箇新LMW-GS基因.GenBank登錄號分彆為GQ892576～GQ892580和GQ892583～GQ892588,其長度為903～1 081 bp.其中,GQ892576、GQ892585、GQ892586、GQ892587和GQ892588具有完整編碼區,可分彆編碼305、351、298、351和307箇氨基痠殘基的成熟蛋白.而GQ892577、GQ892578、GQ892579、GQ892580、GQ892583和GQ892584由于在編碼區內齣現提前終止密碼子,推測為假基因.推導的氨基痠序列結果顯示:它們都具有LMW-m型低分子量穀蛋白亞基的典型結構特徵.
위료알굴화유효이용협253중적우량기인,근거LMW-GS편마기인5'화3'단적보수서렬설계인물,채용PCR방법이협253적기인조DNA위모판진행확증,분별회수병극륭도재체상,측서후득도11개신LMW-GS기인.GenBank등록호분별위GQ892576～GQ892580화GQ892583～GQ892588,기장도위903～1 081 bp.기중,GQ892576、GQ892585、GQ892586、GQ892587화GQ892588구유완정편마구,가분별편마305、351、298、351화307개안기산잔기적성숙단백.이GQ892577、GQ892578、GQ892579、GQ892580、GQ892583화GQ892584유우재편마구내출현제전종지밀마자,추측위가기인.추도적안기산서렬결과현시:타문도구유LMW-m형저분자량곡단백아기적전형결구특정.
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.
