[Cloning and analysis of LMW-GS genes from Triticum aestevum ssp. tibetanum Shao].

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. coil 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.