Abstract
The cDNA encoding a full-length murine immunoglobulin γ1 heavy chain with its native leader sequence, transmembrane and intracellular domains was introduced into transgenic plants. Transformed plants expressed the recombinant polypeptide, but, in contrast to plants expressing the heavy chain without transmembrane sequence, the protein appeared to be associated with a plant cell membrane. Extraction of the membrane-associated heavy chain required the presence of a non-ionic detergent, and immunofluorescence studies of protoplasts demonstrated surface expression of membrane Ig heavy chain on up to 40% of the cells from a transgenic leaf. In plants expressing both the membrane Ig heavy chain and its partner light chain, functional antibody was also localised to the plant cell membrane and retention of the heavy chain at this site appeared to have no effect on the efficiency of antibody assembly. This approach of localising and accumulating recombinant antibody in cell membranes may have a number of applications, including passive immunisation against plant pathogens.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Artsaenko, O., Peisker, M., zur Nieden, U., Fiedler, U., Weiler, E.W., Muntz, K. and Conrad, U. 1995. Expression of a single chain Fv antibody against abscisic acid creates a wilty phenotype in transgenic tobacco. Plant J. 8: 745–750.
Barnes, W.M. 1990. Variable patterns of expression of luciferase in transgenic tobacco leaves. Proc. Natl. Acad. Sci. USA 87: 9183–9187.
Benfey, P.N. and Chua, N.-H. 1990. The cauliflower mosaic virus 35S promoter: combinatorial regulation of transcription in plants. Science 250: 959–966.
Chomczynski, P. and Sacchi, N. 1987. Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162: 156–195.
Firek, S., Draper, J., Owen, M.R.L., Gandecha, A., Cockburn, B. and Whitelam, G.C. 1993. Secretion of a functional single-chain Fv protein in transgenic tobacco plants and cell suspension Cultures. Plant Mol. Biol. 23: 861–870.
Fraley, R.T., Rogers, S.G., Horsch, R.B., Sanders, P.R., Flick, J.S., Adams, S.P., Bittner, M.L., Brand, L.A., Fink, C.L., Fry, J.S., Galluppi, G.R., Goldberg, S.B., Hoffmann, N.L. and Woo, S.C. 1983. Expression of bacterial genes in plant cells. Proc. Natl. Acad. Sci. USA 80: 4803–4807.
Frigerio, L., Vine, N.D., Pedrazzini, E., Hein, M.B., Wang, F., Ma, J.K.-C. and Vitale, A. 2000. Assembly, secretion and vacuolar delivery of a hybrid immunoglobulin in plants. Plant Physiol. 123: 1483–1493.
Galbraith, D.W. 1989. Analysis of higher plants by flow cytometry and cell sorting. Int. Rev. Cytol. 116: 165–228.
Hein, M.B., Tang, Y., McLeod, D.A., Janda, K.D. and Hiatt, A.C. 1991. Evaluation of immunoglobulins from plant cells. Biotechnol. Prog. 7: 455–461.
Hiatt, A.C., Cafferkey, R. and Bowdish, K. 1989. Production of antibodies in transgenic plants. Nature 342: 76–78.
Horsch, R.B., Fry, J.E., Hoffmann, N.L., Eichholtz, D., Rogers, S.G. and Fraley, R.T. 1985. A simple and general method for transferring genes into plants. Science 227: 1229–1231.
Kabat, E.A., Wu, T.T., Perry, H.M., Gottesman, K.S. and Foeller, C. 1999. Sequences of proteins of immunological interest. National Institutes of Health, USA.
Ma, J.K.-C. and Hiatt, K. 1996. Expressing antibodies in plants for immunotherapy. In: M.R.L. Owen and J. Pen (Eds.) Transgenic Plants: A Production System for Industrial and Pharmaceutical Protein, John Wiley, Chichester, UK.
Ma, J.K.-C., Lehner, T., Stabila, P., Fux, C.I. and Hiatt, A. 1994. Assembly of monoclonal antibodies with IgG1 and IgA heavy chain domains in transgenic tobacco plants. Eur. J. Immunol. 24: 131–138.
Owen, M., Gandecha, A., Cockburn, B. and Whitelam, G. 1992. Synthesis of a functional anti-phytochrome single chain Fv protein in transgenic tobacco. Bio/technology 10: 790–794.
Rogers, S.G., Klee, H.J., Horsch, R.B. and Fraley, R.T. 1987. Improved vectors for plant transformation: expression cassette vectors and new selectable markers. Meth. Enzymol. 153: 253–276.
Schouten, A., Roosien, J., van Engelen, F.A., de Jong, G.A.M., Borst-Vrenssen, A.W.M., Zilverentant, J.F., Bosch, D., Stiekema, W.J., Gommers, F.J., Schots, A. and Bakker, J. 1996. The C-terminal KDEL sequence increases the expression level of a single-chain antibody designed to be targeted to both the cytosol and the secretory pathway in transgenic tobacco. Plant Mol. Biol. 30: 781–93.
Smith, R. and Lehner, T. 1989. Characterisation of monoclonal antibodies to common protein epitopes on the cell surface of Strep-tococcus mutans and Streptococcus sobrinus. Oral Microbiol. Immunol. 4: 153–158.
Tavladoraki, P., Benvenuto, E., Trinca, S., De Martinis, D., Cat-taneo, A. and Galeffi, P. 1993. Transgenic plants expressing a functional single-chain Fv antibody are specifically protected from virus attack. Nature 366: 469–472.
Tyler, B.M., Cowman, A.F., Gerondakis, S.D., Adams, J.M. and Bernard, O. 1982. mRNA for surface immunoglobulin gamma chains encodes a highly conserved transmembrane sequence and a 28 residue intracellular domain. Proc. Natl. Acad. Sci. USA 79: 2008–2112.
Yamawaki-Kataoka, Y., Nakai, S., Miyata, T. and Honjo, T. 1982. Nucleotide sequences of gene segments encoding membrane domains of immunoglobulin gamma chains. Proc. Natl. Acad. Sci. USA 79: 2623–2627.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Vine, N.D., Drake, P., Hiatt, A. et al. Assembly and plasma membrane targeting of recombinant immunoglobulin chains in plants with a murine immunoglobulin transmembrane sequence. Plant Mol Biol 45, 159–168 (2001). https://doi.org/10.1023/A:1006477231006
Issue Date:
DOI: https://doi.org/10.1023/A:1006477231006