臺灣博碩士論文加值系統

稻米(Oryza sativa L. )是亞洲國家重要的主食之ㄧ，然稻米胚乳中14-16 kDa的鹽溶性蛋白質，能與有過敏病患之IgE抗體產生交叉反應(cross-reactive)，造成食物過敏反應，甚至休克。由於國內對食物過敏之動物模式研究較少，因此，本研究擬利用藥物誘發動物，以建立動物過敏模式。以腹腔注射途徑對Sprague-Dawley (SD)品系大鼠及Balb/c品系小鼠投予50-1000 μg含佐劑的卵白蛋白(ovalbumin, OVA)建立過敏反應的誘發模式，再以已建立Balb/c小鼠動物模式投予30-120 μg台農67號稻米(Tainung 67, TNG 67)的14-16 kDa蛋白及5、25 μg OVA進行安全性評估，經二次誘導後，第14天解剖取得血清進行分析。研究亦對純化的14-16 kDa過敏蛋白與OVA進行人工胃液及腸液的消化穩定性試驗。粗萃取水稻鹽溶性蛋白質後，再經膠體過濾層析管柱 (column)(Sephadex G-50)純化14-16 kDa蛋白，分離出A、B、C、D四區段，以十二烷基磺酸鈉-聚丙烯醯胺膠電泳(SDS polyacrylamide gel electrophoresis, SDS-PAGE)檢測及利用西方墨點法(Western blot)以過敏病患之血清IgE抗體進行活性檢測，並進行蛋白質質譜鑑定。試驗結果顯示，投予50 μg OVA/佐劑無法明顯誘導SD大鼠產生IgE抗體濃度(35.3 ng/ml)；而Balb/c小鼠投予50-1000 μg OVA/佐劑皆能誘發動物體產生顯著性的IgE抗體濃度(分別為257.5 ng/ml、254.6 ng/ml及256.8 ng/ml)及特異性IgE抗體力價，而投予30、60及120 μg的14-16 kDa過敏蛋白無法明顯誘導Balb/c小鼠產生IgE抗體濃度(分別為236.8 ng/ml、211.0 ng/ml及189.5 ng/ml)，卻可檢測出顯著性的抗原特異性IgE抗體力價。另外，OVA分別可在胃液及腸液中穩定5-60分鐘及2-5分鐘，而稻米14-16 kDa過敏蛋白則是分別在5-15分鐘及0-30秒內，結果顯示稻米14-16 kDa過敏蛋白在胃液及腸液中具穩定。稻米過敏蛋白的研究方面，經SDS-PAGE及Western blot檢測下，B段尖峰具很強烈的活性反應，質譜鑑定下顯示與已知的Rice allergen 16 (RA16)過敏蛋白具相似性。綜合本試驗研究顯示，利用OVA過敏誘導物可成功誘發Balb/c小鼠產生過敏反應，且稻米過敏蛋白14-16 kDa在腹腔途徑投予30-120 μg能明顯產生抗原特異性IgE抗體力價。

Rice (Oryza sativa L.) is an important staple food for Asian. The salt dissolving protein of 14-16 kDa, can react (cross-reactive) with the IgE antibody with allergic patient in the endosperm of rice, cause the allergic response of food, even shock. It is seldom on domestic animal model in evaluating allergic effect to the food. We plan to establish an allergic animal model induced by medicines. The first step, we use SD (Sprague-Dawley) rat and Balb/c mice for allergic animal model test. The allergic effect was induced by injecting 50-1000ug OVA (ovalbumin) on rat and mice. The second step, we assessed the safety of 14-16 kDa protein of Tainung 67 of rice and 5 and 25 μg OVA after challenge in Balb/c mice by exposed. Mice were sacrificed after 14 days treatment through two times drug induction. Serum samples were collected and analyze from mice. The third step, the stability tests of simulated gastric fluid and simulated intestinal fluid to allergic protein of 14-16 kDa of the purification and OVA was investigated. The fraction A, B, C and D from 14-16 kDa allergic protein in rice were separated and collected by gel filtration chromatography column. The protein was pooled separately by SDS polyacrylamide gel electrophoresis (SDS-PAGE) and the allergic activity of rice protein was measured by analyzing the reactivity with the allergic patient’s serum IgE antibody in western blot method. The protein also was analyzed by mass spectrometry. Determination of IgE concentraction (35.3 ng/ml) were not significantly increased after antigen challenge in SD rat exposed to 50 μg OVA with adjuvant. SD rat seems not be caused an allergic response. The IgE concentraction of Balb/c mice were detected in 257.5, 254.6 and 256.8 ng/ml, respectively, after dosing 50-1000ug OVA. The OVA-specific IgE titer was significantly increased as compared with control. Balb/c mice seem to be a suitable allergic animal model. The allergic protein of 14-16 kDa of 30, 60 and 120 ug were all unable to lead the Balb/c mice to produce the IgE antibody concentraction (value is 236.8 ng/ml, 211.0 ng/ml and 189.5 ng/ml respectively), however, food-specific IgE titer were significantly increased after antigen challenge. In addition, OVA was degraded at 5-60 min and 2-5 min respectively among gastric fluid (SGF) and intestinal fluid (SIF), but the rice of 14-16 kDa allergic protein to stable 5-15 minutes and 0-30 second, the result shows that the rice of 14-16 kDa allergic protein have potential anaphylaxis risk to the human health, can stable in gastric fluid and intestinal fluid. In conclusion, this study demonstrated that OVA could induce allergic effect on Balb/c mice. However, 120 μg 14-16 kDa of allergic protein in rice unable to induce IgE antibody significant increased on Balb/c mice by intraperitoneal.

總目錄

中文摘要……………………………………………………………………....I
Abstract…………………………………………………………………….....III
致謝…………………………………………………………………………...V
總目錄………………………………………………………………………..VI
表目錄………………………………………………………………………...IX
圖目錄………………………………………………………………………....X
第一章 前言………………………………………………………………….1
第二章 文獻回顧…………………………………………………………….3
第一節 基因改造生物(GMO)之概論……………………………………….3
一、基因改造生物之定義……………………………………………….3
二、基因改造產品的發展與利用……………………………………….3
三、基因改造生物之爭議……………………………………………….5
四、基因改造生物之安全評估………………………………………….5
第二節 稻米的介紹……………………………………………………….6
一、稻米的組成成份…………………………………………………….6
二、稻米14-16 kDa過敏蛋白的介紹…………………………………..7
第三節 食物過敏的介紹………………………………………………….8
一、食物過敏反應之機制……………………………………………….8
二、食物過敏反應對人體之危害……………………………………….9
第四節 過敏物質在動物模式的建立近況..……………………….…….10
第三章 材料與方法…………………………………………………………16
第一節 材料………………………………………………………………16
一、樣品…………………………………………………………………16
二、動物…………………………………………………………………16
三、藥品…………………………………………………………………16
四、材料…………………………………………………………………18
五、儀器…………………………………………………………………19
第二節 方法………………………………………………………………20
一、動物過敏陽性對照組之建立………………………………………20
1. SD品系大鼠過敏陽性對照組之建立……………………………..20
2. Balb/c品系小鼠過敏陽性對照組之建立………………………….22
2.1. IgG抗體濃度測定……………………………………………....22
2.2. IgE抗體濃度測定………………………………………………23
2.3. IL-4細胞激素濃度測定………………………………………...23
2.4. Passive cutaneous anaphylaxis (PCA)試驗設計……………......24
二、鹽溶性蛋白萃取……………………………………………………24
三、蛋白質定量檢測……………………………………………………25
四、台農67號水稻14-16 kDa分子量測定...…………………………26
1. SDS-PAGE之操作程序…………………………………………....26
1.1. SDS-PAGE膠片的製作…………………………………………26
1.2. SDS-PAGE蛋白質分子量測定………………………………...26
1.3. 膠片乾燥法……………………………………………………..27
五、蛋白質純化………………………………………………………....27
1. 管柱製備……………………………………………………………27 1.1. 管柱清洗與趕氣………………………………………………..27
1.2. 管柱裝填………………………………………………………..28 2. 膠體過濾色層分析(gel filtration chromatography)之純化……….28
六、過敏反應活性測定………………………………………………....29
七、蛋白質質譜鑑定……………………………………………………29
八、胃液消化穩定性(simulated gastric fluid, SGF)之檢測……………30
九、腸液消化穩定性(simulated intestinal fluid, SIF)之檢測…………..30
十、過敏蛋白之動物安全性評估………………………………………31
十一、統計分析…………………………………………………………31
第四章 結果……………………………………………................................32
第一節 動物過敏陽性對照組之建立……………………………………32
一、SD品系大鼠對OVA的過敏反應…………………………………32
二、Balb/c品系小鼠對OVA的過敏反應……………………………..33
三、Balb/c品系小鼠對OVA之PCA試驗……………………………34
第二節 稻米14-16 kDa過敏蛋白的萃取與純化……………………….34
一、台農67號水稻鹽溶性蛋白質的粗萃取…………………………..34
二、台農67號水稻14-16 kDa過敏蛋白的純化……………………...34
第三節 過敏反應活性檢測與鑑定………………………………………35
第四節 過敏蛋白胃液消化穩定反應……………………………………36
第五節 過敏蛋白腸液消化穩定反應……………………………………36
第六節 過敏蛋白之動物安全性評估……………………………………37
一、Balb/c品系小鼠對稻米過敏蛋白及OVA的過敏反應…………..37
二、Balb/c品系小鼠對稻米過敏蛋白及OVA之PCA試驗…………38
第五章 討論…………………………………………………………………39
第六章 結論…………………………………………………........................47
參考文獻………………………………………………………………............48
表……………………………………………………………………................57
圖………………………………………………………………………………59
附錄……………………………………………………………………............78

表目錄

表一、主要穀類過敏原資料表………………………………………………13
表二、質譜值於NCBI資料庫搜尋比對之結果……………………………57
表三、稻米14-16 kDa過敏蛋白部分純化表……………………………….58















圖目錄

圖一、過敏反應誘發途徑……………………………………………………14
圖二、基因改造產品的過敏安全性評估樹狀圖……………………………15
圖三、台農67號水稻照片…………………………………………………59
圖四、Sprague-Dawley品系雌性大鼠血清IgG抗體濃度檢測……………60
圖五、Sprague-Dawley品系雌性大鼠血清IgE抗體濃度檢測……………61
圖六、Sprague-Dawley品系雌性大鼠血清中IL-4細胞激素濃度檢測…...62
圖七、Balb/c品系雌性小鼠血清IgG抗體濃度檢測……………………....63
圖八、Balb/c品系雌性小鼠血清IgE抗體濃度檢測……………………….64
圖九、PCA試驗反應情形……………………………………….……..……65
圖十、Balb/c品系小鼠過敏陽性對照組之PCA試驗結果…………….…..66
圖十一、粗萃取台農67號水稻蛋白分子量之電泳圖.……………….……67
圖十二、台農67號水稻粗蛋白經膠體過濾吸光值分析結果………….…68
圖十三、台農67號水稻粗蛋白經Sephadex G-50純化後之電泳圖………69
圖十四、西方墨點法進行台農67號水稻純化之活性檢測………………70
圖十五、蛋白質質譜分析結果……………………………………………71
圖十六、胃液消化穩定性(simulated gastric fluid, SGF)之檢測…………72
圖十七、腸液消化穩定性(simulated intestinal fluid, SIF)之檢測…….……73
圖十八、過敏蛋白對Balb/c品系雌性小鼠血清IgG抗體濃度檢測……74
圖十九、過敏蛋白對Balb/c品系雌性小鼠血清IgE抗體濃度檢測………75
圖二十、Balb/c品系小鼠對稻米過敏蛋白之PCA試驗結果………….…..76
圖二十一、西方墨點法進行OVA活性檢測………………………..…..…..77

參考文獻

牛惠之、郭華仁、幐沛倫、彭英泰、陳詩欣。2005。基因改造產品-發展、爭議、管理與規範。農委會動植物檢疫防治局。9-37 pp
李國欽、徐慈鴻。2004。GMO/GMF 風險評估與風險管理辦法。基因改造議題-從紛爭到展望。82-99 pp
徐慈鴻、李貽華、李國欽。2003。基轉植物之生物安全性評估及管理。行政院農業委員會農業藥物毒物試驗所技術專刊第70期126號。1-27 pp
張敬宜。2002。綠豆澱粉分支酵素的鑑定。中國醫藥學院營養研究所碩士論文。45-47 pp
郭華仁、周桂田。2004。基改作物的全球經驗。基因改造議題-從紛爭到展望。120-157 pp
陳柏文。2003。乳鐵蛋白之抗菌功能及其在乳房癌檢測之應用。國立中興大學獸醫學系博士論文。162-171 pp
陳樺翰。2004。探討不同碾白度米粒中細胞壁多醣物質特性及其對澱粉理化性質之影響。國立中興大學食品科學系博士論文。2-10 pp
黃三光、曾經洲。2002。基因改造的優勢與潛藏危機。行政院農業委員會農業藥物毒物試驗所技術專刊第63期110號。1-11 pp
潘子明。2006。全球基因改造作物之產量現況與台灣對基因改造產品之管理。科技發展政策報導。341-365 pp
蔡三福、張敬宜、黃振聲、王順成。2005。基因改造產品的毒性與過敏性之安全評估。行政院農業委員會農業藥物毒物試驗所技術專刊第77期134號。1-13 pp
蔡三福、張敬宜、廖俊旺、黃振聲、何素鵬、王順成。2003。白殭菌粗抗原對大鼠之過敏性反應。植物保護協會會刊 45 : 285-294
Akiyama, H., Teshima, R., Sakushima, J. I., Okunuki H., Goda, Y., Sawada, J. I., Toyoda, M., 2001. Examination of oral sensitization with ovalbumin in Brown Norway rats and three strains of mice. Immunology Letter. 78 : 1-5
Alvarez, A. M., Adachi, T., Nakase, M., Aoki, N., Nakamura, R., Matsuda, T., 1995. Classification of rice allergenic protein cDNAs belonging to the α-amylase /trypsin inhibitor gene family. Protein Structure and Molecular Enzymology. 1251 : 201-204
Amano, M., Ogawa, H., Kojima, K., Kamidaira, T., Suetsugu, S., Yoshikawa, M., Satoh, T., Samejima, T., Matsumoto, I., 1998. Identification of the major allergens in wheat flour responsible for baker’s asthma. The Biochemical Journal. 330 : 1229-1234
Armentia, A., Sanchez-Monge, R., Gomez, L., Barber, D., Salcedo, G., 1993. In vivo allergenic activities of eleven purified members of a major allergen family from wheat and barley flour. Clinical and Experimental Allergy. 23 : 410-415
Atkinson, H. A. C., Johnson, I. T., Gee, J. M., Grigoriadou, F., Miller, K., 1996. Brown Norway rat model of food allergy: effect of plant components on the development of oral sensitization. Food and Chemical Toxicology. 34 : 27-32
Birmingham, N., Payankaulam, S., Thanesvorakul, S., Stefura, B., HayGlass, K., Gangur, V., 2003. An ELISA-based method for measurement of food-specific IgE antibody in mouse serum: an alternative to the passive cutaneous anaphylaxis assay. Journal of Immunological Methods. 273 : 89-98
Chang, H. S., Bae, Y.K., Lim, S.K., Jeong, T.C., Kim, H.S., Chung, S.T., Kim, D.S., Nam, D.H., 2001. Allergenicity test of genetically modified soybean in Sprague Dawley rats. Archives Pharmacology Research. 24 : 256-261
Codina, R., Lockey, R. F., Rama, R., 1997. Purification and characterization of a soybean hull allergen responsible for the Barcelona asthma outbreaks. II. Purification and sequencing of the Gly m 2 allergen. Clinical and Experimental Allergy. 27 : 424-430
Dawson, R. M. C., Elliott, D. C.,Elliott, W. H., Jones, K. M., 1969. Data for biochemical research. 2 nd ed. Oxford Univ. Press, London.
Dearman, R. J., Caddick, H., Stone, S., Basketter, D. A., Kimber, I., 2001. Characterization of antibody responses induced in rodents by exposure to food proteins: influence of route of exposure. Toxicology. 167 : 217-231
Dearman, R. J., Kimber, I., 2001. Determination of protein allergenicity: studies in mice. Toxicology Letters. 120 : 181-186
Dearman, R. J., Skinner, R. A., Herouet, C., Labay, K., Debruyne, E., Kimber, I., 2003. Induction of IgE antibody responses by protein allergens: inter-laboratory comparisons. Food and Chemical Toxicology. 41 : 1509-1516
Duijn, G.V., Biert, R.V., Bleeker-Marcelis, H., Peppelman, H., Hessing M., 1999. Detection methods for genetically modified crops. Food Control. 10 : 375-378
FAO/WHO, 2001. Evaluation of allergenicity of genetically modified foods. Allergenicity of genetically modified foods. Report of a joint FAO/WHO Expert Consultation on Allergenicity of Foods Derived from Biotechnology. 22-25 January 2001. Rome, Italy. (www.who.int.)
Figueredo, E., Quirce, S., del Amo, A., Ceresta, J., Arrieta, I., Lahoz, C., Sastre, J., 1999. Beer-induced anaphylaxis: Identification of allergens. Allergy 54 : 630-634
Franken, J., Stephan, U., Meyer, H. E., Konig, W., 1994. Identification of alpha-amylase inhibitor as a major allergen of wheat flour. International Archives of Allergy and Immunology. 104 : 171-174
Fu, T. J., Abbott, U. R., Hatzos, C., 2002. Digestibility of food allerges and nonallergenic proteins in simulated gastric fluid and simulated intestinal fluid-a comparative study. Journal of Agricultural and Food Chemistry. 50 : 7154-7160
Gendel, S. M., 1998. Sequence databases for assessing the potential allergenicity of protein used in transgenic foods. Advances in Food and Nutrition Research. 42 : 63-92
Gonzalez, R., Polo, F., Zapatero, L., Caravaca, F., Carreira, J., 1992. Purification and characterization of major in halant allergens from soybean hulls. Clinical and Experimental Allergy. 22 : 748-755
Hilton, J., Dearman, R. J., Sattar, N., Basketter, D. A., Kimber, I., 1997. Characteristics of antibody responses induced in mice by protein allergens. Food and Chemical Toxicology. 35 : 1209-1218
Hubner, P., Studer, E., Luthy, J., 1999. Quantitation competitive PCR for the detection of genetically modified organisms in food. Food Control. 10 : 353-358
Ikezawa, Z., Tsubaki, K., Yokota, S., 1994. Effect of hypoallergenic wheat (HAW-A1) on atopic dermatitis (AD) with wheat allergy, and its antigenic analysis using sera from patients with AD. Allergy. 43 : 679-688
Jaenisch, R., 1998. Transgenic aminals. Science. 240 : 1468-1474
James, J. M., Sixbey, J. P., Helm, R. M., Bannon, G. A., Burks, A. W., 1997. Wheat α-amylase inhibitor: A second route of allergic sensitization. The Journal of Allergy and Clinical Immunology. 99 : 239-244
Kim, N. I., Kim, W. C., Lee, S. M., Lee, H. K., Rhee, H. I., Choi, Y. S., Jung, Y. H., Cha, S. H., 1999. Isolation of rice allergenic cDNA clones from a rice cDNA library by immunoscreening with a polyclonal antibody specific to 16 kD rice allergenic protein. Experimental and Molecular Medicine. 31 : 185-190
Kimber, I., Dearman, R. J., 2001. Food allergy: what are the issues? Toxicology Letters. 120 : 165-170
Knippels, L. M. J., Penninks, A. H., Spanhaak, S., Houben, G. F., 1998. Oral sensitization to food proteins: a Brown Norway rat model. Clinical and Experimental allergy. 28 : 368-375
Kimoto, M., Yoshikawa, M., Takahashi, K., Bando, N., Okita, M., Tsuji, H., 1998. Identification of allergen in cereals and their hypoallergenization. I. Screening of allergens in wheat and identification of an allergen, Tri a Bd 17K. Annual Report Interdisciple Research Institute Environmental Science. 17 : 53-60
Ladies, G. S. Holsapple, M. P., Astwood, J. D., Kimber, I., Kimber, I., 2003. Approaches to the assessment of the allergenic potential of food from genetically modified crops. Toxicological Sciences. 73 : 8-16
Lehrer, S. B., Reese, G., Malo, J. L., Lahoud, C., Leong-Kee, S., Goldberg, B., Carle, T.,Ebisawa, M., 1999. Corn allergens: IgE antibody reactivity and cross-reactivity with rice, soy, and peanut. International Archives of Allergy and Immunology. 118 : 298-299
Lewkowich, I. P., Rempel, J. D., HayGlass, K. T., 2004. In vivo IgE levels in exogenous antigen stimulated responses: measurement of total IgE as a valid, simple surrogate for Ag-specific IgE. Journal of Immunological Methods. 286 : 123-132
Li, X. M., Serebrisky, D., Lee, S. Y., Huang, C. K., Bardina, L., Schofield, B. H., Stanley, J. S., Burks, A. W., Bannon, G. A., Sampson, H. A., 2000. A murine model of peanut anaphylaxis: T- and B-cell responses to a major peanut allergen mimic human responses. The Journal of Allergy and Clinical Immunology. 106 : 150-158
Matsuda, T., Nomura, R., Sugiyama, M., Nakamura, R., 1991. Immunochemical studies on rice allergenic proteins. Agricultural and Biological Chemistry. 55 : 509-513
Matsuda, T., Sugiyama, M., Nakamura, R., Torii, S., 1988. Purification and properties of an allergenic protenic protein in rice grain. Agricultural and Biological Chemistry. 52 : 1465-1470
Momma, K., Hashimoto, W., Yoon, H. J., Ozawa, S., Fukuda, Y., Kawai, S., Takaiwa, F., Utsumi, S., Murata, K., 2000. Safety assessment of rice genetically modified with soybean glycinin by feeding studies on rats. Bioscience, Biotechnology and Biochemistry. 64 : 1881-1886
Moroz, L. A., Yang, W. H., Kunitz, 1980. A specific allergen in food anaphylaxis. The New England Journal of Medicine. 302 : 1126-1128
Nakamura, R., Matsuda, T., 1996. Rice allergenic protein and molecular-genetic approach for hypoallergenic rice. Bioscience, Biotechnology, and Biochemistry. 60 : 1215-1221
Nakase, M., Adachi, T., Urisu, A., Miyashita, T., Alvarez, A. M., Nagasaka, S., Aoki, N., Nakamura, R., Matsuda, T., 1996. Rice (Oryza sativa L.) α-amylase inhibitors of 14-16 kDa are potential allergens and products of a multigene family. Journal of Agricultural and Food Chemistry. 44 : 2624-2628
Nakase, M., Usui, Y., Alvarez-Nakase, A. M., Adachi, T., Urisu, A., Nakamura, R., Aoki, N., Kitajima, K., Matsuda, T., 1998. Cereal allergens: rice-seed allergens with structural similarity to wheat and barley allergens. Allergy. 53 : 55-57
Ogawa, T., Babdo, N., Tsuji, H., Nishikawa, K., Kitamura, K., 1995. α-subunit of β-conglycinin, an allergenic protein recognized by IgE antibodies of soybean-sensitive patients with atopic dermatitis. Bioscience, Biotechnology, and Biochemistry. 59 : 831-833
Ogawa, T., Babdo, N., Tsuji, H., Okajima, H., Nishikawa, K., Sasaoka, K., 1991. Investigation of the IgE-binding proteins in soybean by immunoblotting with the sera of the soybean sensitive patients with atopic dermatitis. Journal of Nutritional Science and Vitaminology. 37 : 555-565
Ogawa, T., Tsuji, H., Babdo, N., Kitamura, K., Zhu, Y. I., Hirano, H., Nishikawa, K., 1993. Identification of the soybean allergenic protein, Gly m Bb 30K, with the soybean seed 34-kDa Oil-body-associated protein. Bioscience, Biotechnology, and Biochemistry. 57 : 1030-1033
Ogawa, T., Tsuji, H., Bando, N., Sasaoka, K., 1992. Identification of a soybean protein producing IgE antibodies in BALB/c Mice. Bioscience, Biotechnology, and Biochemistry. 56 : 978-979
Pilegaard, K., Madsen, C., 2004. An oral Brown Norway rat model for food allergy: comparison of age, sex, dosing volume, and allergen preparation. Toxicology. 196 : 247-257
Rihs, H. P., Chen, Z., Rueff, F., Petersen, A., Rozynek, P., Heiman, H., Baur, X., 1999. IgE binding of the recombinant allergen soybean profiling is mediated by conformational epitopes. The Journal of Allergy and Clinical Immunology. 104 : 1293-1301
Sanchez-Monge, R., Gomez, L., Barber, D., Lopez-Otin, C., Armentia, A., Salcedo, G., 1992. Wheat and barley allergens associated with baker’s asthma. Glycosylated subunits of the a-amylase inhibitor family have enhanced IgE-binding capacity. The Biochemical Journal. 281 : 401-405
Shibasaki, M., Suzuki, S., Nemoto, H., Kuroume, T., 1979. Allergenicity and lymphocyte-stimulating property of rice protein. The Journal of Allergy and Clinical Immunology. 64 : 259-265
Tada, Y., Nakase, M., Adachi, T., Nakamura, R., Shimada, H., Takahashi, M., Fulimura, T., Matsuda, T., 1996. Reduction of 14-16 kDa allergenic proteins in transgenic rice plants by antisense gene. FEBS Letters. 391 : 341-345
Talbot, W., 1990. Manufacture of biopharmaceutical proteins by mammalian cell culture system. Biotechnology Advances. 8 : 729-739
Taylor, S., Lehrer, S. B., 1996. Priniciples and characteristics of food allergens. Critical Reviews in Food Science and Nutrition, Allergenicity of Foods Produced by Genetic Modification. 36 : S91-S118
Thomas, K., Aalbers, M., Bannon, G. A., Bartels, M., Dearman, R. J., Esdaile, D. J., Fu, T. J., Glatt, C. M., Hadfield, N., Hatzos, C., Hefle, S. L., Heylings, J. R., Goodman, R. E., Henry, B., Herouet, C., Holsapple, M., Ladics, G. S., Landry, T. D., Macintosh, S. C., Rice, E. A., Privalle, L. S., Steiner, H. Y., Teshima, R., Ree, R. V., Woolhiser, M., Zawodny, J., 2004. A multi-laboratory evaluation of a common in vitor pepsin digestion assay protocol used in assessing the safety of novel proteins. Regulatory Toxicology and Pharmacology. 39 : 87-98
Tsuji, H., Babdo, N., Hiemori, Kitamura, K., Zhu, Y. I., Hirano, H., Nishikawa, K., Ogawa, T., 1997. Purification and characterization of soybean allergen Gly m Bb 28K. Bioscience, Biotechnology, and Biochemistry. 61 : 942-947
Tsuji, H., Kimoto, M., Natori, Y., 2001. Allergens in major crops. Nutrition Research. 21 : 925-934
Urisu, A., Yamada, Y., Masuda, S., Komada, H., Wada, E., Kondo, Y., Horiba, F., Tsuruta, M., Yasaki, T., Yamada, M., Torii, S., Nakamura, R., 1991. 16-kDa rice protein is one of the major allergens in rice grain extract and responsible for cross-allergenicity between cereal grains in the Poaceae family. International Archives of Allergy and Immunology. 96 : 244-252
Watanabe, M., Miyakawa, J., Ikezawa, Z., Suzuki, Y., Hirao, T., Yoshizawa, T., Arai, S., 1990. Production of hypoallergenic rice by enzymatic decomposition of constituent proteins. Journal of Food Science. 55 : 781-783