C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 9/90 (2006.01) C07H 21/04 (2006.01) C12N 1/21 (2006.01) C12N 15/74 (2006.01) C12P 21/06 (2006.01)
Patent
CA 2597647
This invention relates to methods of expressing eukaryotic proteins in prokaryotic hosts, particularly eukaryotic proteins that require formation of disulfide bridges for biological activity. Various approaches are used including fusion to thioredoxin, cytoplasmic expression of disulfide isomerases, deficiencies in thioredoxin and/or glutathione reductases, deficiencies in proteases, and the like. The method is applicable to express monomelic and dimeric forms of the eukaryotic protein with biological activity such as monomeric and dimeric forms of a disintegrin or a disintegrin domain. Included are the vectors, host cells expressing the proteins, the expressed proteins and methods of using the proteins.
La présente invention concerne des procédés d~expression de protéines eucaryotes dans des hôtes procaryotes, en particulier des protéines eucaryotes qui exigent la formation de ponts disulfure pour une activité biologique. On utilise diverses approches comprenant la fusion en thiorédoxine, l'expression cytoplasmique de disulfure-isomérases, des insuffisances en thiorédoxine et/ou en glutathione-réductases, des insuffisances en protéases et analogues. On peut utiliser le procédé pour exprimer des formes monomères et dimères de la protéine eucaryote avec une activité biologique telle que des formes monomères et dimères d'une disintégrine ou d'un domaine d~une disintégrine. La présente invention comprend les vecteurs, les cellules hôtes exprimant les protéines, les protéines exprimées et les procédés utilisant les protéines.
Markland Frances
Minea Radu
Gowling Lafleur Henderson Llp
Pivotal Biosciences Inc.
University Of Southern California
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