Molecular Cloning and Nucleotide Sequences of Heavy and Light Chain Genes of a Monoclonal Antibody to Potato Virus Y Coat Protein

Kazusato OHSHIMA; Akihiro MATSUURA; Takaaki NAKAYA; Eishiro SHIKATA; Kokichi KIKUCHI; Ikuo KIMURA

doi:10.3186/jjphytopath.60.600

Kazusato OHSHIMA, Akihiro MATSUURA, Takaaki NAKAYA, Eishiro SHIKATA, Kokichi KIKUCHI, Ikuo KIMURA

Author information

Keywords: monoclonal antibody gene, heavy and light chains, nucleotide sequence, potato virus Y

JOURNAL FREE ACCESS

1994 Volume 60 Issue 5 Pages 600-607

DOI https://doi.org/10.3186/jjphytopath.60.600

Details

Published: October 25, 1994 Received: May 23, 1994 Available on J-STAGE: April 03, 2009 Accepted: - Advance online publication: - Revised: -
Correction information
Date of correction: April 03, 2009 Reason for correction: - Correction: CITATION Details: Wrong : 3. Bird, R.E., Hardman, K.D., Jacobson, J.W., Johnson, S., Kaufman, B.M., Lee, S.-M., Lee, T., Pope, S.H., Riordan, G.S. and Whitlow, M. (1988). Single-chain antigen-binding proteins. Science 242: 423-426.
4. Buchner, J. and Rudolph, R. (1991). Renaturation, purification and characterization of recombinant Fab-fragments produced in Escherichia coli. Bio/Technology 9: 157-162.
5. Chiang, Y.L., Sheng-Dong, R., Brow, M.A. and Larrick, J.W. (1989). Direct cDNA cloning of the rearranged immunoglobulin variable region. Bio Techniques 7: 360-366.
6. De Neve, M., De Loose, M., Jacobs, A., Van Houdt, H., Kaluza, B., Weidle, U., Van Montagu, M. and Depicker, A. (1993). Assembly of an antibody and its derived antibody fragment in Nicotiana and Arabidopsis. Trans. Res. 2: 227-237.
7. De Sutter, K., Feys, V., Van de Voorde, A. and Fiers, W. (1992). Production of functionally active murine and murine:: human chimeric F (ab')₂ fragments in COS-1 cells. Gene 113: 223-230.
8. Düring, K., Hippe, S., Kreuzaler, F. and Schell, J. (1990). Synthesis and self-assembly of a functional monoclonal antibody in transgenic Nicotiana tabacum. Plant Mol. Biol. 15: 281-293.
9). Feinberg, A.P. and Vogelstein, B. (1984). A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 137: 260-267.
10). Glisin, V., Crkvenjakov, R. and Byus, C. (1974). Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry 13: 2633-2637.
11. Grunstein, M. and Hogness, D.S. (1975). Colony hybridization: A method for the isolation of cloned DNAs that contain a specific gene. Proc. Natl. Acad. Sci. USA 72: 3961-3965.
12. Gubler, U. and Hoffman, B.J. (1983). A simple and very efficient method for generating cDNA libraries. Gene 25: 263-269.
13. Halk, E.L. and De Boer, S.H. (1985). Monoclonal antibodies in plant-disease research. Annu. Rev. Phytopath. 23: 321-350.
15. Hataya, T., Inoue, A.K., Ohshima, K., Shikata, E. (1994). Characterization and strain identification of a potato virus Y isolate non-reactive with monoclonal antibodies specific to the ordinary and necrotic strains. Intervirology 37: 12-19.
16. Hiatt, A. and Ma, J.K.-C. (1992). Monoclonal antibody engineering in plants. FEBS Lett. 307: 71-75.
17. Hiatt, A. and Cafferkey, R. and Bowdish, K. (1989). Production of antibodies in transgenic plants. Nature 342: 76-78.
18. Horwitz, A.H., Chang, C.P., Better, M., Hellstrom, K.E. and Robinson, R.R. (1988). Secretion of functional antibody and Fab fragment from yeast cells. Proc. Natl. Acad. Sci. USA 85: 8678-8682.
19. Huse, W.D., Sastry, L., Iverson, S.A., Kang, A.S., Alting-Mees, M., Burton, D.R., Benkovic, S.J. and Lerner, R.A. (1989). Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda. Science 246: 1275-1281.
21. Kabat, E.A., Wu, T.T., Bilofsky, H., Reid-Miller, M. and Perry, H. (eds.) (1983). Sequences of proteins of immunological interest. U.S. Department of Health and Human Services.
22. Köhler, G. and Milstein, C. (1975). Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256: 495-497.
23. Kofler, R., Strohal, R., Balderas, R.S., Johnson, M.E., Noonan, D.J., Duchosal, M.A., Dixon, F.J. and Theofilopoulos, A.N. (1988). Immunogloublin kappa-chain variable region gene complex organization and immunoglobulin genes encoding anti-DNA autoantibodies in lupus mice. J. Clin. Invest. 82: 852-860.
24. Komaromy, M., Clayton, L., Rogers, J., Robertson, S., Kettman, J. and Wall, R. (1983). The structure of the mouse immunoglobulin in γ₃ membrane gene segment. Nucl. Acids Res. 11: 6775-6785.
25. Larrick, J.W., Danielsson, L., Brenner, C.A., Wallace, E.F., Abrahamson, M., Fry, K.E. and Borrebaeck, A.K. (1989). Polymerase chain reaction using mixed primers: Cloning of human monoclonal antibody variable region genes from single hybridoma cells. Bio/Technology 7: 934-938.
26. McCafferty, J., Griffiths, A.D., Winter, G. and Chiswell, D.J. (1990). Phage antibodies: filamentous phage displaying antibody variable domains. Nature 348: 552-554.
27. Ohshima, K. (1995). Monoclonal antibodies: Viruses. In Molecular Methods in Plant Pathology (Singh, R.P. and Singh, U.P. eds.). CRC Press, Boca Raton, Florida (in press).
28. Ohshima, K. and Shikata, E. (1990). On the screening procedures of ELISA for monoclonal antibodies against three luteoviruses. Ann. Phytopath Soc. Japan 56: 219-228.
29. Ohshima, K., Hataya, T., Sano, T., Inoue, A.K. and Shikata, E. (1991). Comparison of biological properties, serological characteristics and amino acid sequences of coat protein between potato virus Y ordinary strain and necrotic strain. Ann. Phytopath. Soc. Japan 57: 615-622.
30. Owen, M., Gandecha, A., Cockburn, B. and Whitelam, G. (1992). Synthesis of a functional antiphytochrome single-chain Fv protein in transgenic tobacco. Bio/Technology 10: 790-794.
31. Plückthun, A. (1991). Antibody engineering: advances from the use of Escherichia coli expression systems. Bio/Technology 9: 545-551.
32. Saiki, R.K., Gelfand, D.H., Stoffel, S., Scharf, S.J., Higuchi, R., Horn, G.T., Mullis, K.B. and Erlich, H.A. (1988). Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 230: 1350-1354.
33. Sakano, H., Maki, R., Kurosawa, Y., Roeder, W. and Tonegawa, S. (1980). Two types of somatic recombination are necessary for the generation of complete immunoglobulin heavy-chain genes. Nature 286: 676-683.
34. Sander, E. and Dietzgen, R.G. (1984). Monoclonal antibodies against plant viruses. Adv. Virus Res. 29: 131-168.
35. Sanger, F., Nicklen, S. and Coulsen, A.R. (1977). DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74: 5463-5467.
36. Tavladoraki, P., Benvenuto, E., Trinca, S., De Martinis, D., Cattaneo, A. and Galeffi, P. (1993). Transgenic plants expressing a functional single-chain Fv antibody are specifically protected from virus attack. Nature 366: 469-472.
37. Towbin, H., Staehelin, T. and Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and applications. Proc. Natl. Acad. Sci. USA 76: 4350-4354.
38. Ullrich, A., Shine, J., Chirgwin, J., Pictet, R., Tischer, E., Rutter, W.J. and Goodman, H.M. (1977). Rat insulin genes: Construction of plasmids containing the coding sequences. Science 196: 1313-1318.
39. Ward, E.S., Güssow, D., Griffiths, A.D., Jones, P.T. and Winter, G. (1989). Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli. Nature 341: 544-546.
40. Weidle, U.H., Borgya, A., Mattes, R., Lenz, H. and Buckel, P. (1987). Reconstitution of functionally active antibody directed against creatine kinase from separately expressed heavy and light chains in non-lymphoid cells. Gene 51: 21-29.
41. Weissenhorn, W., Weiss, E., Schwirzke, M., Kaluza, B. and Weidle, U.H. (1991). Chimerization of antibodies by isolation of rearranged genomic variable regions by the polymerase chain reaction. Gene 106: 273-277.
42. Wood, C.R., Boss, M.A., Kenten, J.H., Calvert, J.E., Roberts, N.A. and Emtage, J.S. (1985). The synthesis and in vivo assembly of functional antibodies in yeast. Nature 314: 446-449.

Right : 1. Benton, W.D. and Davis, R.W. (1977). Screening λgt recombinant clones by hybridization to single plaques in situ. Science 196: 180-182.
2. Benvenuto, E., Ordàs, R.J., Tavazza, R., Ancora, G., Biocca, S., Cattaneo, A. and Galeffi, P. (1991). ‘Phytoantibodies’: a general vector for the expression of immunoglobulin domains in transgenic plants. Plant Mol. Biol. 17: 865-874.
3. Bird, R.E., Hardman, K.D., Jacobson, J.W., Johnson, S., Kaufman, B.M., Lee, S.-M., Lee, T., Pope, S.H., Riordan, G.S. and Whitlow, M. (1988). Single-chain antigen-binding proteins. Science 242: 423-426.
4. Buchner, J. and Rudolph, R. (1991). Renaturation, purification and characterization of recombinant Fab-fragments produced in Escherichia coli. Bio/Technology 9: 157-162.
5. Chiang, Y.L., Sheng-Dong, R., Brow, M.A. and Larrick, J.W. (1989). Direct cDNA cloning of the rearranged immunoglobulin variable region. Bio Techniques 7: 360-366.
6. De Neve, M., De Loose, M., Jacobs, A., Van Houdt, H., Kaluza, B., Weidle, U., Van Montagu, M. and Depicker, A. (1993). Assembly of an antibody and its derived antibody fragment in Nicotiana and Arabidopsis. Trans. Res. 2: 227-237.
7. De Sutter, K., Feys, V., Van de Voorde, A. and Fiers, W. (1992). Production of functionally active murine and murine:: human chimeric F (ab')₂ fragments in COS-1 cells. Gene 113: 223-230.
8. Düring, K., Hippe, S., Kreuzaler, F. and Schell, J. (1990). Synthesis and self-assembly of a functional monoclonal antibody in transgenic Nicotiana tabacum. Plant Mol. Biol. 15: 281-293.
9). Feinberg, A.P. and Vogelstein, B. (1984). A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 137: 260-267.
10). Glisin, V., Crkvenjakov, R. and Byus, C. (1974). Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry 13: 2633-2637.
11. Grunstein, M. and Hogness, D.S. (1975). Colony hybridization: A method for the isolation of cloned DNAs that contain a specific gene. Proc. Natl. Acad. Sci. USA 72: 3961-3965.
12. Gubler, U. and Hoffman, B.J. (1983). A simple and very efficient method for generating cDNA libraries. Gene 25: 263-269.
13. Halk, E.L. and De Boer, S.H. (1985). Monoclonal antibodies in plant-disease research. Annu. Rev. Phytopath. 23: 321-350.
14. Hamlyn, P.H., Brownlee, G.G., Cheng, C-C., Gait, M.J. and Milstein, C. (1978). Complete sequence of constant and 3′ noncoding regions of an immunoglobulin mRNA using the dideoxynucleotide method of RNA sequencing. Cell 15: 1067-1075.
15. Hataya, T., Inoue, A.K., Ohshima, K., Shikata, E. (1994). Characterization and strain identification of a potato virus Y isolate non-reactive with monoclonal antibodies specific to the ordinary and necrotic strains. Intervirology 37: 12-19.
16. Hiatt, A. and Ma, J.K.-C. (1992). Monoclonal antibody engineering in plants. FEBS Lett. 307: 71-75.
17. Hiatt, A. and Cafferkey, R. and Bowdish, K. (1989). Production of antibodies in transgenic plants. Nature 342: 76-78.
18. Horwitz, A.H., Chang, C.P., Better, M., Hellstrom, K.E. and Robinson, R.R. (1988). Secretion of functional antibody and Fab fragment from yeast cells. Proc. Natl. Acad. Sci. USA 85: 8678-8682.
19. Huse, W.D., Sastry, L., Iverson, S.A., Kang, A.S., Alting-Mees, M., Burton, D.R., Benkovic, S.J. and Lerner, R.A. (1989). Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda. Science 246: 1275-1281.
20. Jouvin-Marche, E. and Rudikoff, S. (1986). Evolution of a Vκ gene family. Immunogenetics 24: 191-201.
21. Kabat, E.A., Wu, T.T., Bilofsky, H., Reid-Miller, M. and Perry, H. (eds.) (1983). Sequences of proteins of immunological interest. U.S. Department of Health and Human Services.
22. Köhler, G. and Milstein, C. (1975). Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256: 495-497.
23. Kofler, R., Strohal, R., Balderas, R.S., Johnson, M.E., Noonan, D.J., Duchosal, M.A., Dixon, F.J. and Theofilopoulos, A.N. (1988). Immunogloublin kappa-chain variable region gene complex organization and immunoglobulin genes encoding anti-DNA autoantibodies in lupus mice. J. Clin. Invest. 82: 852-860.
24. Komaromy, M., Clayton, L., Rogers, J., Robertson, S., Kettman, J. and Wall, R. (1983). The structure of the mouse immunoglobulin in γ₃ membrane gene segment. Nucl. Acids Res. 11: 6775-6785.
25. Larrick, J.W., Danielsson, L., Brenner, C.A., Wallace, E.F., Abrahamson, M., Fry, K.E. and Borrebaeck, A.K. (1989). Polymerase chain reaction using mixed primers: Cloning of human monoclonal antibody variable region genes from single hybridoma cells. Bio/Technology 7: 934-938.
26. McCafferty, J., Griffiths, A.D., Winter, G. and Chiswell, D.J. (1990). Phage antibodies: filamentous phage displaying antibody variable domains. Nature 348: 552-554.
27. Ohshima, K. (1995). Monoclonal antibodies: Viruses. In Molecular Methods in Plant Pathology (Singh, R.P. and Singh, U.P. eds.). CRC Press, Boca Raton, Florida (in press).
28. Ohshima, K. and Shikata, E. (1990). On the screening procedures of ELISA for monoclonal antibodies against three luteoviruses. Ann. Phytopath Soc. Japan 56: 219-228.
29. Ohshima, K., Hataya, T., Sano, T., Inoue, A.K. and Shikata, E. (1991). Comparison of biological properties, serological characteristics and amino acid sequences of coat protein between potato virus Y ordinary strain and necrotic strain. Ann. Phytopath. Soc. Japan 57: 615-622.
30. Owen, M., Gandecha, A., Cockburn, B. and Whitelam, G. (1992). Synthesis of a functional antiphytochrome single-chain Fv protein in transgenic tobacco. Bio/Technology 10: 790-794.
31. Plückthun, A. (1991). Antibody engineering: advances from the use of Escherichia coli expression systems. Bio/Technology 9: 545-551.
32. Saiki, R.K., Gelfand, D.H., Stoffel, S., Scharf, S.J., Higuchi, R., Horn, G.T., Mullis, K.B. and Erlich, H.A. (1988). Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 230: 1350-1354.
33. Sakano, H., Maki, R., Kurosawa, Y., Roeder, W. and Tonegawa, S. (1980). Two types of somatic recombination are necessary for the generation of complete immunoglobulin heavy-chain genes. Nature 286: 676-683.
34. Sander, E. and Dietzgen, R.G. (1984). Monoclonal antibodies against plant viruses. Adv. Virus Res. 29: 131-168.
35. Sanger, F., Nicklen, S. and Coulsen, A.R. (1977). DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74: 5463-5467.
36. Tavladoraki, P., Benvenuto, E., Trinca, S., De Martinis, D., Cattaneo, A. and Galeffi, P. (1993). Transgenic plants expressing a functional single-chain Fv antibody are specifically protected from virus attack. Nature 366: 469-472.
37. Towbin, H., Staehelin, T. and Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and applications. Proc. Natl. Acad. Sci. USA 76: 4350-4354.
38. Ullrich, A., Shine, J., Chirgwin, J., Pictet, R., Tischer, E., Rutter, W.J. and Goodman, H.M. (1977). Rat insulin genes: Construction of plasmids containing the coding sequences. Science 196: 1313-1318.
39. Ward, E.S., Güssow, D., Griffiths, A.D., Jones, P.T. and Winter, G. (1989). Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli. Nature 341: 544-546.
40. Weidle, U.H., Borgya, A., Mattes, R., Lenz, H. and Buckel, P. (1987). Reconstitution of functionally active antibody directed against creatine kinase from separately expressed heavy and light chains in non-lymphoid cells. Gene 51: 21-29.
41. Weissenhorn, W., Weiss, E., Schwirzke, M., Kaluza, B. and Weidle, U.H. (1991). Chimerization of antibodies by isolation of rearranged genomic variable regions by the polymerase chain reaction. Gene 106: 273-277.
42. Wood, C.R., Boss, M.A., Kenten, J.H., Calvert, J.E., Roberts, N.A. and Emtage, J.S. (1985). The synthesis and in vivo assembly of functional antibodies in yeast. Nature 314: 446-449.

Date of correction: April 03, 2009 Reason for correction: - Correction: PDF FILE Details: -

Abstract

cDNA to a plant virus-monoclonal antibody gene was cloned and sequenced. The source of mRNAs was a hybridoma producing an IgG₃ monoclonal antibody (42C07) which bound to a coat protein of potato virus Y. We have constructed cDNA libraries using a bacteriophage lambda ZAP II vector from the mRNA of the hybridomas. Screening of the libraries with polymerase chain reaction-mediated probes specific to variable and a part of constant regions of the heavy and light chain genes yielded clones of both chains. Numbers of the nucleotides encoding heavy and light chains were 1383 and 717, and numbers of the deduced amino acids were 461 and 239, respectively. Comparisons of the deduced amino acid sequences with those of other immunoglobulin (Ig) previously reported showed that they were full-length of the antibody gene. Furthermore, the heavy and light chains of 42C07 were successfully expressed separately in Escherichia coli as judged by reactivities with anti-mouse Ig antibody.

Corresponding author

Correction information

Register with J-STAGE for free!