Molecular cloning and characterization of a cysteine-rich 16.6-kDa prolamin in rice seeds.
An alcohol-soluble storage protein, a 16.6-kDa prolamin found in rice seeds, was purified from both the total protein body and purified type I protein body fractions. The partial amino acid sequences of three tryptic peptides generated from the purified polypeptide were analyzed. A part of the 16.6-kDa prolamin cDNA was amplified from developing seed mRNA by the reverse transcribed polymerase chain reaction using an oligo (dT) primer and a primer which was synthesized based on the partial amino acid sequence. The amplified product was used to isolate the full-length cDNA clone (lambda RP16) from a developing seed cDNA library. The cDNA has an open reading frame encoding a hydrophobic polypeptide of 149 amino acids. The polypeptide was rich in glutamine (20.0%), cysteine (10.0%), and methionine (6.9%). The cysteine content was higher than those of most other rice storage proteins. Messenger RNA of the 16.6-kDa prolamin was detected in seeds, but not in other aerial tissues.
Mitsukawa
N
N
Laboratory of Genetic Engineering, Faculty of Agriculture Kyoto Prefectural University, Japan.
Konishi
R
R
Uchiki
M
M
Masumura
T
T
Tanaka
K
K
D000595:Amino Acid Sequence; D018554:Avena / Q000235:genetics; D003001:Cloning, Molecular; D003545:Cysteine; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012275:Oryza / Q000235:genetics*; D010446:Peptide Fragments / Q000737:chemistry; D010802:Phylogeny; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics*; D055315:Prolamins; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D003313:Zea mays / Q000235:genetics
D010446:Peptide Fragments; D010940:Plant Proteins; D055315:Prolamins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D003545:Cysteine