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Production and purification of recombinant human interleukin-6 secreted by the yeast Saccharomyces cerevisiae

(1991) EUROPEAN JOURNAL OF BIOCHEMISTRY. 198(1). p.217-222
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Abstract
The coding region of the human interleukin-6 (hIL6) gene was fused to the prepro secretion signal of the alpha-mating factor gene in several yeast host strains. It was found that the KEX-2 protease was unable to cleave the prepro-Lys-Arg-Pro-IL6 sequence, but that unspecific cleavage of the precursor protein had occurred. The prepro-Lys-Arg-Ala-Pro-IL6 sequence, however, was correctly recognized and cleaved by the KEX-2 protease, and IL6 was efficiently secreted into the culture medium. The N-terminal Ala-Pro peptide was removed during processing by wild-type yeast strains, but was retained in a ste13 mutant. IL6 as well as the aberrant proteins were not glycosylated. The transformed cells could secrete up to 30-mu-g/ml IL6. The protein was purified from the medium to homogeneity by ion-exchange chromatography and gel filtration, and had a specific activity of about 2 x 10(8) IU/mg in a proliferation assay.
Keywords
DNA, CELL, TRANSFORMATION, PROTEINS, STRUCTURAL GENE, O-GLYCOSYLATION, ESCHERICHIA-COLI, DIPEPTIDYL AMINOPEPTIDASE, AMINO-ACID SEQUENCE, PREPRO-ALPHA-FACTOR

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Citation

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MLA
Guisez, Yves, Bernard Tison, Joël Vandekerckhove, et al. “Production and Purification of Recombinant Human Interleukin-6 Secreted by the Yeast Saccharomyces Cerevisiae.” EUROPEAN JOURNAL OF BIOCHEMISTRY 198.1 (1991): 217–222. Print.
APA
Guisez, Y., Tison, B., Vandekerckhove, J., Demolder, J., Bauw, G., Haegeman, G., Fiers, W., et al. (1991). Production and purification of recombinant human interleukin-6 secreted by the yeast Saccharomyces cerevisiae. EUROPEAN JOURNAL OF BIOCHEMISTRY, 198(1), 217–222.
Chicago author-date
Guisez, Yves, Bernard Tison, Joël Vandekerckhove, Jan Demolder, Guy Bauw, Guy Haegeman, Walter Fiers, and Roland Contreras. 1991. “Production and Purification of Recombinant Human Interleukin-6 Secreted by the Yeast Saccharomyces Cerevisiae.” European Journal of Biochemistry 198 (1): 217–222.
Chicago author-date (all authors)
Guisez, Yves, Bernard Tison, Joël Vandekerckhove, Jan Demolder, Guy Bauw, Guy Haegeman, Walter Fiers, and Roland Contreras. 1991. “Production and Purification of Recombinant Human Interleukin-6 Secreted by the Yeast Saccharomyces Cerevisiae.” European Journal of Biochemistry 198 (1): 217–222.
Vancouver
1.
Guisez Y, Tison B, Vandekerckhove J, Demolder J, Bauw G, Haegeman G, et al. Production and purification of recombinant human interleukin-6 secreted by the yeast Saccharomyces cerevisiae. EUROPEAN JOURNAL OF BIOCHEMISTRY. 1991;198(1):217–22.
IEEE
[1]
Y. Guisez et al., “Production and purification of recombinant human interleukin-6 secreted by the yeast Saccharomyces cerevisiae,” EUROPEAN JOURNAL OF BIOCHEMISTRY, vol. 198, no. 1, pp. 217–222, 1991.
@article{221548,
  abstract     = {The coding region of the human interleukin-6 (hIL6) gene was fused to the prepro secretion signal of the alpha-mating factor gene in several yeast host strains. It was found that the KEX-2 protease was unable to cleave the prepro-Lys-Arg-Pro-IL6 sequence, but that unspecific cleavage of the precursor protein had occurred. The prepro-Lys-Arg-Ala-Pro-IL6 sequence, however, was correctly recognized and cleaved by the KEX-2 protease, and IL6 was efficiently secreted into the culture medium. The N-terminal Ala-Pro peptide was removed during processing by wild-type yeast strains, but was retained in a ste13 mutant. IL6 as well as the aberrant proteins were not glycosylated. The transformed cells could secrete up to 30-mu-g/ml IL6. The protein was purified from the medium to homogeneity by ion-exchange chromatography and gel filtration, and had a specific activity of about 2 x 10(8) IU/mg in a proliferation assay.},
  author       = {Guisez, Yves and Tison, Bernard and Vandekerckhove, Joël and Demolder, Jan and Bauw, Guy and Haegeman, Guy and Fiers, Walter and Contreras, Roland},
  issn         = {0014-2956},
  journal      = {EUROPEAN JOURNAL OF BIOCHEMISTRY},
  keywords     = {DNA,CELL,TRANSFORMATION,PROTEINS,STRUCTURAL GENE,O-GLYCOSYLATION,ESCHERICHIA-COLI,DIPEPTIDYL AMINOPEPTIDASE,AMINO-ACID SEQUENCE,PREPRO-ALPHA-FACTOR},
  language     = {eng},
  number       = {1},
  pages        = {217--222},
  title        = {Production and purification of recombinant human interleukin-6 secreted by the yeast Saccharomyces cerevisiae},
  volume       = {198},
  year         = {1991},
}