小麦LMW-GS基因的分离与归类  

Identification of New Genes Encoding the Low-Molecular-Weight Glutenins in Wheat

作　　者：崔翠菊[1] 马凤云[1] 王洪文[1] 毛跃建[1] 汪越胜[1] 何光源[1] 

机构地区：[1]华中科技大学中英HUST-RRes基因工程和基因组学联合实验室,武汉430074

出　　处：《分子植物育种》2009年第4期788-792,共5页Molecular Plant Breeding

基　　金：国家自然科学基金项目(30871524)资助

摘　　要：本研究以湖北省小麦栽培品种鄂麦18为材料,采用Glu-D3位点特异性的引物,以叶片DNA为模板,进行PCR扩增。经过PCR产物与克隆载体的连接、连接产物转化感受态E.coliDH5α、蓝白斑筛选以及菌落PCR鉴定,获得34个阳性克隆;经测序分析获得两个LMW-GS基因完整编码序列,在GenBank中的基因登记注册号为DQ822593和DQ822596,其中,DQ822593编码序列全长1287bp,DQ822596编码序列全长908bp,所推导的蛋白质都含有8个半胱氨酸残基,1～19个氨基酸为前导信号肽序列,前者属于LMW-GS中的TypeⅠ类,后者属于TypeⅡ类,目前正在鉴定这两类基因的功能。PCR was used as a rapid altemative method to isolate allelic of the low molecular weight subunit （LMW-GS） genes of wheat. The primers specific to Glu-D3 locus were designed based on published LMW-GS gene sequence data. The PCR products were inserted into pUCm-T vector, the recombinant was transferred into E. coli DH5α, the bacteria underwent blue-white selection and colony PCR identification, and two gene clones were obtained. The size ofgene sequences DQ822593 and DQ822596 were 1 287 bp and 908 bp, respectively, LMW-GS encoded by two genes in wheat contained 8 Cys residues and short signal peptides （the first to nineteenth amino acids）, ranged into LMW-GS Type Ⅰ and Type Ⅱ. These genes could be applied in the gene function research by the gene transformation method.

关 键 词：小麦(Triticum AESTIVUM L.) 低分子量麦谷蛋白 基因分离 

分 类 号：S512.1[农业科学—作物学] Q343.1[生物学—遗传学]