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Research Papers
Bio/Technology  5, 595 - 598 (1987)
doi:10.1038/nbt0687-595

Molecular Cloning and Synthesis of Biologically Active Human Tissue Inhibitor of Metalloproteinases in Yeast

Michel Kaczorek1, Nadine Honoré1, Véronique Ribes1, Pierre Dehoux1, Philippe Cornet1, Terence Cartwright*, 1 & Rolf E. Streeck1

  1Unité des Applications du Génie Génétique (G3), Institut Pasteur, 28 Rue du Docteur Roux, 75724 Paris Cedex 15, France.

  *Laboratoire Roger Bellon, 37260 Monts, France.

Tissue inhibitor of metalloproteinases (TIMP) is a widely distributed glycoprotein that stochiometrically inactivates metalloproteinases involved in connective tissue catabolism. Here we report the cDNA cloning of TIMP from human fibroblastic MRC5 cells using a single 42−base oligonucleotide probe. Expression in S. cerevisiae of complete TIMP cDNA yielded insoluble protein aggregates. Biologically active TIMP was reconstituted from the yeast product by a denaturation/renaturation procedure.

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