Loss of soybean trypsin inhibitor in callus as monitored by inhibition enzyme immunoassay
Tóm tắt
A monoclonal antibody was produced against Kunitz soybean inhibitor (KSBTI) and used in an inhibition enzyme immunoassay (EIA). The inhibition EIA was as sensitive as competetive EIAs and was easily modified for other protein-antibody interactions. The KSBTI assay described detected KSBTI in complex mixtures from 100 μg/ml to 50 ng/ml and did not react with the Bowman-Birk trypsin inhibitor. The assay was used to examine levels of KSBTI inGlycine max hypocotyl-derived callus tissue. The developing hypocotyls contained 0.21 μg KSBTI per mg of fresh tissue. This level of KSBTI rapidly decreased when placed in culture and was undetectable 6 days later. The decrease in KSBTI correlated with the development of callus.
Tài liệu tham khảo
Brandon, D. L.; Bates, A. H.; Friedman, M. Enzyme-linked immunoassay of soybean Kunitz trypsin inhibitor using monoclonal antibodies. J. Food Sci. 53:102–106; 1988.
Gamborg, O. L.; Miller, R. A.; Ojima, K. Nutrient requirements of suspension cutures of soybean root cells. Exp. Cell. Res. 50:148–151; 1968.
Hughes, M. A.; Dunn, M. A. The use of immunochemical techniques to study plant proteins. Plant Mol. Biol. Rep. 3:17–23; 1985.
Hitchcock, C. H. S.; Bailey, F. J.; Crimes, A. A., et al. Determination of soya proteins in food using an enzyme-linked immunosorbent assay procedure. J. Sci. Food Agric. 32:157–165; 1981.
Kerns, H. R.; Barwale, U. B.; Meyer, M. M., et al. Correlation of cotyledonary node shoot proliferation and somatic embryoid development in suspension cultures of soybean (Glycine max L. Merr.) Plant Cell Rep. 5:140–143; 1986.
Liener, I. E. Trypsin inhibitors: concern for human nutrition or not? Nutrition 116:920–924; 1973.
Liener, I. E. Legume toxins in relation to protein digestibility: A review. J. Food Sci. 41:1076–1081; 1976.
Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tabacco tissue cultures. Physiol. Plant 15:473–497; 1962.
Oi, V. T.; Herzenberg, L. A. Imunoglobulin-producing hybrid cell lines. Mishell, B. B.; Shiigi, S. M. eds. Selected methods in cellular immunology. San Francisco: W. H. Freeman and Co.; 1980:351–372.
Rackis, J. J.; Anderson, R. L. Isolation of four soybean trypsin inhibitors by DEAE-cellulose chromatography. Biochem. Biophys. Res. Commun. 15:230–235; 1964.
Ryan, C. A. Proteolytic enzymes and their inhibitors in plants. Ann. Rev. Plant. Physiol. 24:173–196; 1973.
Stahli, C.; Staehelin, T.; Miggiano, V., et al. High frequencies of antigen-specifc hybridomas: dependence on immunization parameters and prediction by spleen cell analysis. J. Immunol. Meth. 32:297–304; 1980.
Tan-Wilson, A. L.; Hartl, P. M.; Delfel, N. E., et al. Differential expression of Kunitz and Bowman-Birk soybean proteinase inhibitors in plant and callus tissue. Plant Physiol. 78:310–314; 1985.
Wilson, K. A. The structure, function and evolution of legume proteinase inhibitors. In: Ory, R. L., ed. Antinutrients and natural toxicants in food. Westport, Connecticut: Food and Nutrition Press; 1981:187–202.