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Identification and isolation of a new x‐type HMW glutenin subunit 1Dx1.6 t gene from Aegilops tauschii
Author(s) -
An X. L.,
Li X. H.,
Xiong X. J.,
Yan Y. M.,
Zhang Y. Z.,
Gao L. Y.,
Wang A. L.,
Wang K.,
Zeller F. J.,
Hsam S. L. K.
Publication year - 2009
Publication title -
plant breeding
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.583
H-Index - 71
eISSN - 1439-0523
pISSN - 0179-9541
DOI - 10.1111/j.1439-0523.2008.01543.x
Subject(s) - glutenin , aegilops tauschii , biology , genetics , gene , protein subunit , coding region , microbiology and biotechnology , open reading frame , homology (biology) , scn3a , sequence analysis , nucleic acid sequence , peptide sequence , g alpha subunit , genome
A new x‐type HMW glutenin subunit, designated as 1Dx1.6 t from Aegilops tauschii was identified and characterized by SDS‐PAGE and MALDI‐TOF‐MS. This subunit is located between 1Dx2 and 1Dx1.5 t and possesses a molecular mass ( M r ) of 88565.8 Da. Its complete coding sequence was amplified via allele‐specific PCR (AS‐PCR), and the amplified product was cloned and sequenced. The authenticity of the cloned 1Dx1.6 t gene was confirmed by successful expression of its open reading frame in Escherichia coli. The molecular characterization of 1Dx1.6 t gene showed that its coding region consisted of 2541 bp encoding a polypeptide of 845 amino acid residues. Sequence comparison to previously characterized 1Dx1.5 t subunit which is related to good dough quality of bread wheat indicated that the 1Dx1.6 t subunit displayed high homology, but possesses 14 residue substitutions and a nonapeptide insertion. A total of 12 single‐nucleotide polymorphisms (1 per 212 bp) was identified in the 1Dx1.6 t allele (11 in repetitive domain and 1 in the C‐terminal domain), which could facilitate the design of AS‐PCR markers.