Identification and isolation of a new x-type HMW glutenin subunit 1Dx1.6t gene from Aegilops tauschii |
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| Authors: | X. L. An X. H. Li X. J. Xiong Y. M. Yan Y. Z. Zhang L. Y. Gao A. L.Wang K. Wang F. J. Zeller S. L. K. Hsam |
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| Affiliation: | Key Laboratory of Genetics and Biotechnology, College of Life Science, Capital Normal University, 1000 Beijing, China;;Chair of Plant Breeding, Technical University of Munich, D-85350 Freising-Weihenstephan, Germany;;Corresponding author, E-mail: |
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| Abstract: | A new x-type HMW glutenin subunit, designated as 1Dx1.6t from Aegilops tauschii was identified and characterized by SDS-PAGE and MALDI-TOF-MS. This subunit is located between 1Dx2 and 1Dx1.5t 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 1Dx1.5t subunit which is related to good dough quality of bread wheat indicated that the 1Dx1.6t 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. |
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| Keywords: | Aegilops tauschii HMW-GS MALDI-TOF-MS molecular cloning |
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