Abstract
We have used a modified CaMV 35S promoter to direct the expression of chimaeric genes encoding human serum albumin (HSA) in transgenic potato and tobacco plants. To secrete the protein, either the human prepro-sequence or the signal sequence from the extracellular tobacco protein PR-S was used. We demonstrate secretion of HSA with both types of signal sequences in transgenic leaf tissue and in suspension cultures. HSA produced in transgenic potato plants was purified to chromatographic homogeneity. N-terminal amino acid sequence analysis revealed that the processing of the precursor protein was dependent on the type of signal sequence. Expression of the human preproHSA gene lead to partial processing of the precursor and secretion of proHSA. Fusion of HSA to the plant PR-S presequence resulted in cleavage of the presequence at its natural site and secretion of correctly processed HSA that is indistinguishable from the authentic human protein.
This is a preview of subscription content, access via your institution
Relevant articles
Open Access articles citing this article.
-
Effect of methyl jasmonate and salycilic acid synergism on enhancement of bilobalide and ginkgolide production by immobilized cell cultures of Ginkgo biloba
Bioresources and Bioprocessing Open Access 19 May 2016
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
References
Gasser, C.S. and Fraley, R.T. 1989. Genetically engineering plants for crop improvement. Science 244: 1293–1299.
Goodman, R.M., Houck, C.M. and Comai, L. 1987. Molecular farming. PCT WO 87/00865.
Vandekerckhove, J., Van Damme, J., Van Lijsebettens, M., Botterman, J., De Block, M., Vandewiele, M., De Clercq, A., Leemans, J., Van Montagu, M. and Krebbers, E. 1989. Enkephalines produced in transgenic plants using modified 2S seed storage proteins. Bio/Technology 7: 929–932.
Hiatt, A., Cafferkey, R. and Bowdish, K. 1989. Production of antibodies in transgenic plants. Nature 342: 76–78.
Judah, J.D., Gamble, M. and Steadman, J.H. 1973. Biosynthesis of serum albumin in rat liver, evidence for the existence of ‘proalbumin’. Biochem. J. 134: 1083–1091.
Brennan, S.O., Owen, M.C., Boswell, D.R., Lewis, J.H. and Carrell, R.W. 1984. Circulating proalbumin associated with a variant proteinase inhibitor. Biochim. Biophys. Acta 802: 24–28.
Etcheverry, T., Forrester, W. and Hitzeman, R. 1986. Regulation of the chelatin promoter during the expression of human serum albumin or yeast phosphoglycerate kinase in yeast. Bio/Technology 4: 726–730.
Latta, M., Knapp, M., Sarmientos, P., Brefort, G., Becquart, J., Guerrier, L., Jung, G. and Mayaux, J.-F. 1987. Synthesis and purification of mature human serum albumin from E. coli. Bio/Technology 5: 1309–1314.
Saunders, C.W., Schmidt, B.J., Mallonee, R.L. and Guyer, M.S. 1987. Secretion of human serum albumin from Bacillus subtilis. J. Bact. 169: 2917–2925.
Chen, J. and Varner, J.E. 1985. An extracellular matrix protein in plants: characterization of a genomic clone for carrot extensin. EMBO J. 4: 2145–2151.
De Loose, M., Alliotte, L., Gheysen, G., Gielen, J., Soetaert, P., Van Montagu, M. and Inze, D. 1988. Primary structure of a hormonally regulated B-glucanase of Nicotiana plumbiginifolia. Gene 70: 13–23.
Hooft Van Huijsduijnen, R.A.M., Cornelissen, B.J.C., Loon, L.C.V., Boom, J.H.V., Tromp, M. and Bol, J.F. 1985. Virus-induced synthesis of messenger RNAs for precursors of pathogenesis-related proteins in tobacco. EMBO J. 4: 2167–2171.
Guilley, H., Dudley, R.K., Jonard, G., Balazs, E. and Richards, K.E. 1982. Transcription of Cauliflower Mosaic Virus DNA: detection of promoter sequences and characterization of transcripts. Cell 30: 763–773.
Kay, R., Chan, A., Dayly, M. and McPherson, J. 1987. Duplication of CaMV 35S promoter sequences creates a strong enhancer for plant genes. Science 236: 1299–1302.
Brederode, F.T., Koper-Zwarthoff, E.C. and Bol, J.F. 1980. Complete nucleotide sequence of alfalfa mosaic virus RNA 4. Nucleic Acids Res. 8: 2213–2223.
Jobling, S.A. and Gehrke, L. 1987. Enhanced translation of chimaeric messenger RNAs containing a plant viral untranslated leader sequence. Nature 325: 622–625.
Lawn, R.M., Adelman, J., Bock, S.C., Franke, A.E., Houck, C.M., Najarian, C., Seeburg, P.H. and Wion, K.L. 1981. The sequence of human serum albumin cDNA and its expression in E. coli. Nucleic Acid Res. 9: 6103–6114.
Ditta, G., Stanfield, S., Corbiu, D. and Helinski, D. 1980. Broad host range DNA cloning system for gram-negative bacteria: construction of a genebank of Rhizobium meliloti. Proc. Nat. Aca. Sci. USA 77: 7347–7351.
Hoekema, A., Huisman, M.J., Molendijk, L., Van den Elzen, P.J.M. and Cornelissen, B.J.C. 1989. The genetic engineering of two commercial potato cultivars for resistance to potato virus X. Bio/Technology 7: 273–278.
Reiss, B., Sprengel, R., Will, H. and Schaller, H. 1984. A new and sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene 30: 217–223.
Hendriks, T., Van den Berg, B.M. and Schram, A.W. 1985. Cellular location of peroxidase isoenzymes in leaf tissue of Petunia and their affinity for Concanavalin A-Sepharose. Planta 164: 89–95.
Persson, B., Flinta, C., Heijne, G.V. and Jornvall, H. 1985. Structures of N-terminally acetylated proteins. Eur. J. Biochem. 152: 523–527.
Von Heijne, G. 1986. A new method for predicting signal sequence cleavage sites. Nucl. Acid Res. 14: 4683–4690.
Miernyk, J.A. and Nelsen, T.C. 1989. Patterns of amino acids near the signal-sequence cleavage sites of plant secretory proteins. Plant Physiol. 89: S–62.
Ryan, C.A. and Walker-Simmons, M. 1981. Plant Proteinases. p. 321–350. In: The Biochemistry of Plants, Vol. 6. Stumpf, P.K. and Conn, E.E. (Eds.). Academic Press, New York.
Brennan, S.O. and Carrell, R.W. 1978. A circulating variant of human proalbumin. Nature 274: 908–909.
Appleton, D.W. and Sarkar, B. 1971. The absence of specific copper(II) binding site in dog albumin. J. Biol. Chem. 246: 5040–5046.
Lau, S. and Sarkar, B. 1971. Ternary coordination complex between human serum albumin, copper(II) and L-histidine. J. Biol. Chem. 246: 5938–5943.
Wetzler, H.C., Mignery, G.A., Fisher, L.M. and Park, W.D. 1989. Analysis of a chimeric class-I patatin-GUS gene in transgenic potato plants: high-level expression in tubers and sucrose-inducible expression in cultured leaf and stem explants. Plant Mol. Biol. 12: 41–50.
Baulcombe, D.C., Saunders, G.R., Bevan, M.W., Mayo, M.A. and Harrison, B.D. 1986. Expression of biologically active viral satellite RNA from the nuclear genome of transformed plants. Nature 321: 446–449.
Yanisch-Perron, C., Vieira, J. and Messing, J. 1985. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors. Gene 33: 103–119.
Sleate, D.E., Gallie, D.R., Jefferson, R.A., Bevan, M.W., Turner, P.C. and Wilson, T.M.A. 1987. Characterization of the 5′-leader sequence of tobacco mosaic virus RNA as a general enhancer of translation in vitro. Gene 217: 217–225.
Bevan, M. 1984. Agrobacterium vectors for plant transformation. Nucleic Acids Res. 22: 8711–8721.
Cornelissen, B.J.C. Hooft van Huijsduijnen, R.A.M. and Bol, J.F. 1986. A tobacco mosaic virus-induced tobacco protein is homologous to the sweet-tasting protein thaumatin. Nature 321: 531.
Kunkel, T.A. 1985. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc. Nat. Acad. Sci. USA 82: 488–492.
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.
Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497.
Verwoerd, T.C., Dekker, B.M.M. and Hoekema, A. 1989. A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res. 6: 2362.
Krieg, P.A. and Melton, D.A. 1984. Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nucleic Acids Res. 12: 7057–7070.
Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–254.
Szewcyk, B. and Summers, D.F. 1988. Preparative elution of proteins blotted to Immobilon membranes. Anal. Biochem. 168: 48–53.
Giulian, G.G., Moss, R.L. and Greaser, M. 1983. Improved methodology for analysis and quantitation of proteins on one-dimensional silver-stained slab gels. Anal. Biochem. 129: 277–287.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sijmons, P., Dekker, B., Schrammeijer, B. et al. Production of Correctly Processed Human Serum Albumin in Transgenic Plants. Nat Biotechnol 8, 217–221 (1990). https://doi.org/10.1038/nbt0390-217
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nbt0390-217
This article is cited by
-
Nicotiana hairy roots for recombinant protein expression, where to start? A systematic review
Molecular Biology Reports (2023)
-
The Journey of 1-Keto-1,2,3,4-Tetrahydrocarbazole Based Fluorophores: From Inception to Implementation
Journal of Fluorescence (2022)
-
Optimization of inside and outside factors to improve recombinant protein yield in plant
Plant Cell, Tissue and Organ Culture (PCTOC) (2017)
-
Effect of methyl jasmonate and salycilic acid synergism on enhancement of bilobalide and ginkgolide production by immobilized cell cultures of Ginkgo biloba
Bioresources and Bioprocessing (2016)