Exp Mol Med.  1999 Dec;31(4):185-190.

Isolation of rice allergenic cDNA clones from a rice cDNA library by immunoscreening with a polyclonal antibody specific to 16 kD rice allergenic protein

• Kim NI
• Kim WC
• Lee SM
• Lee HK
• Rhee HI
• Choi YS
• Jung YH
• Cha SH

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.

Keyword

rice; allergy; allergen; cDNA library

MeSH Terms

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