[Cloning of low-molecular-weight glutenin subunit gene from Sichuan wheat landrace AS1643 and its secondary structure prediction].
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%.
Chen
Hua-Ping
HP
College of Biology and Science, Sichuan Agricultural University, Ya'an 625014, China.
Huang
Qian-Ming
QM
Wei
Yu-Ming
YM
Zheng
You-Liang
YL
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D005983:Glutens / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D014908:Triticum / Q000235:genetics*
D010940:Plant Proteins; D005983:Glutens; C037302:glutenin