Molecular Cloning And Sequence Analysis Of Low-molecular-weight Glutenin Subunit Genes From Yumai 34

The low-molecular-weight glutenin subunit(LMW-GS)of wheat has significant influence on dough processing quality. It represents about 1/3 of the amount in seed storage proteins and 60% of glutenin. The LMW-GSs are encoded by Glu-A3, Glu-B3 and Glu-D3 on the short arm of chromosomes 1A, 1B and 1D, respectively. Cloning and identification a novel LMW-GS gene and studying its function are of great significance. In the present paper,a novel LMW-GS gene, LMWY34-1 (GenBank No. GU183486), and a pseudogene,LMWY34-2(GenBank No. HM131885) ,were amplified and cloned with a pair of primers of LMW-GS genes from the wheat Variety Yumai 34. The main results were as follows:(1)A pair of primers of LMW-GS genes was used to amplify and clone LMW-GS genes from the wheat Variety Yumai 34. One PCR band with about 900bp in length was detected on 1% agarose gel. The length was nearly equal to the coding region of known LMW-GS genes;(2)After positive clone screen and sequencing two LMW-GS genes, designated as LMWY34-1and LMWY34-2, were obtained from the wheat Variety Yumai 34 .The GenBank accession numbers are GU183486 and HM131885, respectively. The LMWY34-1 is a typical LMW-GS gene. The whole coding region of this gene was 906 bp, encoding a protein of 302 amino acid residues. But LMWY34-2 is a pseudogene due to the presence of an inframe stop codon ;(3)The estimated molecular weight of LMWY34-1is about 32kDa, indicating that it is C type of LMW-GS. Comparisons of amino acid sequences showed that it had typical structure of LMW-m type in N-terminal region. The LMWY34-1 had high sequence similarity with the reported LMW-GS genes encoded by the GluD3-4 locus.