摘要
根据小麦低分子量谷蛋白基因序列保守区设计的引物W12 2 7/ 12 2 8对特异种质资源 西藏半野生小麦(Triticumaestivumssp .tibetanumShao)AS90 8总DNA进行PCR扩增 ,得到约 95 0bp的DNA片段 ,分离纯化后连接到pBluescriptSK(+/ )T载体上 ,转化、筛选后选取 3种不同类型的阳性克隆进行测序 ,获得 3个不同的基因序列 ,Ti bet1、Tibet2和Tibet3。其中 ,Tibet1(GenBank登录号 :AY2 994 5 7)、Tibet2 (GenBank登录号 :AY2 994 5 8)的编码区长度分别为 10 4 1bp和 90 6bp ,可编码 32 6和 2 81个氨基酸残基的成熟蛋白。Tibet3(GenBank登录号 :AY2 994 5 9)由于编码区内有 2个提前终止密码子而为不可编码成熟蛋白的假基因。序列比较显示AY2 994 5 7、AY2 994 5 8和AY2 994 5 9分别与GenBank中Glu D3、Glu A3和Glu A3位点编码的LMW GS基因AY2 14 4 5 0、AJ2 930 99和AB0 6 2 871有较高的一致性 ,且序列结构非常相似 ,因而推测它们与之有类似的品质功能 。
The full coding regions (open reading frame,ORF) of LMW-GS genes were amplified from Triticum aestevum ssp.tibetanum Shao accession AS908 by using the primer pairs W1227 and W1228,which were designed according to wheat LMW-GS conservative domains.The amplified DNA fragments were separated and recovered from agrose gel,subsequently ligated into pBluescript SK(+/-)-T vector,and then transformed into E.coli strain DH10B.Three positive clones were screened out and sequenced.Among which,Tibet1 (GenBank accession No:AY299457) and Tibet2 (GenBank accession No:AY299458) are two expressed LMW glutenin genes.Their coding regions are 1 041 bp and 901 bp,and encode two mature proteins with 326 and 281 amino acid residues,respectively.Tibet3 (GenBank accession No:AY299459) is a pseudogene due to that two stop codons are in its coding region.Squence multipule alignment analysis suggested that Tibet1,Tibet2 and Tibet3 have the higher similarity in their structure and quality functions with the known LMW-GS genes with GenBank accession No:AY214450,AJ293099 and AB062871 encoded by Glu-D3,Glu-A 3 and Glu-A3,respectively.
基金
国家 8 63计划项目 (编号 :2 0 0 3AA2 0 710 0 )
全国优秀博士学位论文作者专项基金资助项目 (编号 :2 0 0 3 5 7) ~~~
