Gene structure and expression of rice seed allergenic proteins belonging to the ?-amylase/trypsin inhibitor family

Adachi, Takahiro; Izumi, Hidehiko; Yamada, Takehisa; Tanaka, Kunisuke; Takeuchi, Shunji; Nakamura, Ryô; Matsuda, Tsukasa

doi:10.1007/bf00019940

Cited by 96 publications

(78 citation statements)

References 35 publications

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“…To express antisense allergen RNA in transgenic rice seeds, a 0.55 kb cDNa for 16 kDa allergen (RA17) [9] and the promoters of four rice-seed specific genes, the rice allergen gene (RAG1) [15], branching enzyme I gene (BE), prolamin gene and glutelin gene, were used. The 1.0 kb rice allergen promoter and the 0.8 kb glutelin promoter [16] were PCR-amplified by using the genomic clone, pRAG1 [15], and the rice genomic DNA prepared from rice leaves as a template, respectively.…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…The 1.0 kb rice allergen promoter and the 0.8 kb glutelin promoter [16] were PCR-amplified by using the genomic clone, pRAG1 [15], and the rice genomic DNA prepared from rice leaves as a template, respectively. The 0.7 kb prolamin promoter and the 1.6 kb BE promoter were excised by EcoRI-SmaI digestion from a cloned prolamin gene, PG5a, [17] and by BamHI-SmaI digestion from a cloned BE gene [18], respectively.…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…The subsequent screening of the immunologically cross-reacting proteins has demonstrated that most of the 14-16 kDa proteins in seed salt-soluble proteins are cross-reactive with the monoclonal antibody, 25B9 [7], and also reactive with the patients' serum IgE (unpublished data). The sequence data obtained so far on the cDNAs encoding the 14-16 kDa proteins showed that the 14-16 kDa allergen genes belong to a multigene family [8,15]. These results indicate that to produce 'hypo-allergenic rice' the expression of all genes encoding these homologous proteins must be suppressed.…”

Section: Y Tada Et Al/febs Letters 391 (1996) 341-345mentioning

confidence: 99%

“…In the present study, however, the rice-derived gene promoters including the allergen gene itself were used for the antisense gene expression in transgenic rice plants, since rice grains harvested from transgenic rice plants containing only rice-derived genes are most suitable for transgenic food from a view point of public acceptance. The Y-flanking region of RAG1 [15] was used at first as a promoter to transcribe the antisense gene, but the reduction of the allergenic proteins in transgenic rice plants was insufficient and unstable (unpublished data). Therefore, to increase the antisense gene expression in transgenic rice seeds, some other promoters of riceseed specific genes, prolamin [17], glutelin [16] and starch branching enzyme [18] were used in combination with RA17 gene promoter.…”

Section: Y Tada Et Al/febs Letters 391 (1996) 341-345mentioning

confidence: 99%

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Reduction of 14–16 kDa allergenic proteins in transgenic rice plants by antisense gene

et al. 1996

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An antisense gene strategy was applied to suppress the 14-16 kDa allergen gene expression in maturing rice seeds. Gene constructs producing antisense RNAs of the 16 kDa allergen under the control of some rice seed-specific promoters were introduced into rice by electroporation. Immunoblot and RNA blot analyses of the seeds from the transgenic rice plants using the allergen-specific monoclonal antibody and a sequencespecific antisense RNA probe demonstrated that the 14-16 kDa allergen proteins and their transcripts of the seeds from several transgenic lines were present in much lower in amounts than those of the seeds from parental wild-type rice. The high levels of reduction observed were stably inherited in at least three generations.Key words: Rice allergen; Transgenic rice; Antisense RNA has been used experimentally to inhibit gene expression in bacteria, yeast, plant and animal cells [12], and the antisense strategy has also been reported to be practically applicable to transgenic crop plants, to which genes that produce antisense RNA are introduced to suppress the gene expression [13,14]. In the present study, the antisense genes for the 16 kDa allergen were introduced into rice by electroporation, and specific suppression of the 14-16 kDa allergen gene expression, resulting in the reduction of corresponding allergenic proteins in the mature seeds, was examined for regenerated transgenic rice plants and their progeny. The results show that antisense RNA markedly reduced the mRNA and protein contents of the 14-16 kDa allergens in the transgenic rice seeds. Materials and methods

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Section: Plasmid Constructionmentioning

confidence: 99%

Section: Plasmid Constructionmentioning

confidence: 99%

Section: Y Tada Et Al/febs Letters 391 (1996) 341-345mentioning

confidence: 99%

Section: Y Tada Et Al/febs Letters 391 (1996) 341-345mentioning

confidence: 99%

See 2 more Smart Citations

Reduction of 14–16 kDa allergenic proteins in transgenic rice plants by antisense gene

et al. 1996

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“…A cDNA clone of rice allergen, RA17, was isolated from a rice (Oryza sativa L. var japonica cv Nipponbare) cDNA library using oligonucleotide probe, and turned out to be an α-amylase/trypsin inhibitor family (Izumi et al, 1982). Other rice allergen (RA) cDNA clones have been identified by screening rice genomic or cDNA libraries by way of nucleic acid hybridization method using a radioactive-labeled RA17 DNA fragment (Adachi et al, 1993;Alvarez et al, 1995). In this study, we generated a rice cDNA library from Korean rice cultivar using λ UniZap vector.…”

Section: Introductionmentioning

confidence: 99%

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

Kim²,

Lee³

et al. 1999

Exp Mol Med

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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 λ λ λ λ 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.

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Use of enzymes in the processing of protein products from rice bran and rice flour

Shih¹,

Champagne

Daigle

et al. 1999

Nahrung

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Gene structure and expression of rice seed allergenic proteins belonging to the ?-amylase/trypsin inhibitor family

Cited by 96 publications

References 35 publications

Reduction of 14–16 kDa allergenic proteins in transgenic rice plants by antisense gene

Reduction of 14–16 kDa allergenic proteins in transgenic rice plants by antisense gene

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

Use of enzymes in the processing of protein products from rice bran and rice flour

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