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HEREDITAS ›› 2007, Vol. 29 ›› Issue (7): 859-866.doi: 10.1360/yc-007-0859

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Cloning of low-molecular-weight glutenin subunit gene from Sichuan wheat landrace AS1643 and its secondary structure prediction

CHEN Hua-Ping1,2,3, HUANG Qian-Ming1, WEI Yu-Ming2,3, ZHENG You-Liang 2,3   

  1. 1. College of Biology and Science, Sichuan Agricultural University, Ya,an 625014, China;
    2. Ministry of Education for Crop Genetic Resources and Improvement in Southwest China, Sichuan Agricultural University, Ya,an 625014, China;
    3. Triticeae Research Institute, Sichuan Agricultural University, Dujiangyan 611830 China
  • Received:2007-02-09 Revised:2007-04-29 Online:2007-07-10 Published:2007-07-10
  • Contact: CHEN Hua-Ping

Abstract: The full-length coding region (open reading frame, ORF) of low-molecular-weight glutenin subunit (LMW-GS ) gene was amplified from Sichuan wheat landrace accession AS1643 by using the primer pairs P1/P2, which were designed according to wheat LMW-GS conservative domains. The amplified DNA fragment was separated and recovered from agarose gel, subsequently cloned into pMD18-T vector, and then transformed into E. coli strain DH5a. One positive clone LMW-AS1643 (GenBank accession No. EF 190322) was selected and sequenced. It had a coding region of 909 bp, and encoded a mature protein of 302 amino acid residues. Sequence analysis suggested that LMW-AS1643 was a typical LMW-GS gene. The deduced amino acid sequence comparison suggested that LMW-AS1643 had a high similarity with other known LMW-GS genes, with the highest similarity of 93.40 %Bioinformatics analysis indicated that among the three types of secondary structures, content of coil was the highest with 67.90 %, a-helix was the second with 30.46%, and b-sheet was the lowest with 1.64%.

Key words: structure prediction, gene cloning, low-molecular-weight glutenin subunit, wheat landrace