Four rice seed cDNA clones belonging to the alpha-amylase/trypsin inhibitor gene family encode potential rice allergens.
Four rice seed proteins encoded by cDNAs belonging to the alpha-amylase/trypsin inhibitor gene family were overexpressed as TrpE-fusion proteins in E. coli. The expressed rice proteins were detected by SDS-PAGE as major proteins in bacterial cell lysates. Western blot analyses showed that all the recombinant proteins were immunologically reactive to rabbit polyclonal antibodies and to a mouse monoclonal antibody (25B9) specific for a previously isolated rice allergen of 16 kDa. Some truncated proteins from deletion mutants of the cDNAs retained their reactivity to the specific antibodies. These results suggest that the cDNAs encode potential rice allergens and that some epitopes of the recombinant proteins are still immunoreactive when they are expressed as their fragments.
Alvarez
A M
AM
Department of Applied Biological Sciences, School of Agricultural Sciences, Nagoya University, Japan.
Fukuhara
E
E
Nakase
M
M
Adachi
T
T
Aoki
N
N
Nakamura
R
R
Matsuda
T
T
D000485:Allergens / Q000737:chemistry*; D015153:Blotting, Western; D018076:DNA, Complementary / Q000737:chemistry*; D018744:DNA, Plant / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D012275:Oryza / Q000378:metabolism*; D012639:Seeds / Q000737:chemistry*; D009626:Terminology as Topic; D014361:Trypsin Inhibitors / Q000235:genetics*; D000516:alpha-Amylases / Q000235:genetics*
D000485:Allergens; D018076:DNA, Complementary; D018744:DNA, Plant; D014361:Trypsin Inhibitors; D000516:alpha-Amylases