Isolation of rice allergenic cDNA clones from a rice cDNA library by immunoscreening with a polyclonal antibody specific to 16 kD rice allergenic protein.
- Author:
Nam il KIM
1
;
Woo Chang KIM
;
Sang Mi LEE
;
Hee Kyeong LEE
;
Hae Ik RHEE
;
Yong Soon CHOI
;
Yeon Ho JUNG
;
Sang Hoon CHA
Author Information
1. Kangwon National University, College of Agriculture & Life Sciences, Division of Food Sci. & Biotechnology, Chunchon Korea.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
rice;
allergy;
allergen;
cDNA library
- MeSH:
Allergens/immunology;
Allergens/genetics*;
Amino Acid Sequence;
Animal;
Antibodies/immunology;
Antibody Specificity;
Cloning, Molecular;
Cross Reactions;
DNA, Complementary;
Female;
Gene Library;
Genetic Vectors;
Immunoblotting/methods*;
Molecular Sequence Data;
Plant Proteins/immunology;
Plant Proteins/genetics*;
Rats;
Rats, Inbred Strains;
Rice/genetics*;
Sequence Analysis, DNA
- From:Experimental & Molecular Medicine
1999;31(4):185-190
- CountryRepublic of Korea
- Language:English
-
Abstract:
Clinical cases of type-1 hypersensitive reaction to rice (Oryza sativa) have been reported in western countries as well as in Japan. Among rice proteins, 14-16 kD globulin proteins encoded by multiple gene family have been identified as major rice allergens. In this study, a rice cDNA library was constructed using lambda UniZap vector and screened with a rat anti-16 kD globulin protein polyclonal antibody in order to isolate Korean rice allergenic cDNA clones. Five independent cDNA clones, termed RAK1-5, were obtained after second rounds of plaque assay and immunoblot analysis. These clones encoded 13-19 kD recombinant proteins upon IPTG induction, which were identified by the polyclonal antibody in immunoblot analysis. DNA sequencing analysis showed that RAK1-4 have 99% sequence homology with RA5b, and RAK5 is closely related with RA14c. This result indicated that RA5b gene is widely distributed in our cDNA library among other possible rice allergenic genes, and more study is needed to isolate heterogeneous or novel rice allergen genes. Copyright 2000 Academic Press.
