Recombinant proteinase from clostridium hystolyticum and the...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/57 (2006.01) C12N 9/52 (2006.01) C12N 5/06 (2006.01) C12N 5/08 (2006.01)

Patent

CA 2208324

A process is proposed for breaking down cellular tissue and releasing the constituent cells or cell groups by incubating the cellular tissue with a recombinant neutral protease from Clostridium hystolyticum. The protease in question is coded by (a) a DNA of the nucleotide 1027-1965 from SEQ ID NO:3 or by a DNA complementary to it; (b) nucleic acids which hybridise with a DNA of the nucleotide 1027-1965 from SEQ ID NO:3; (c) nucleic acids which would hybridise with one of the nucleic acids mentioned in (a) or (b) without degeneration of the genetic code. The protease is a product of a prokaryotic or eukaryotic expression of an exogenic nucleic acid. Incubation continues until the release of cells or cell groups in the desired quantity and separation of the cells or cell groups from the cellular tissue fractions.

L'invention concerne un procédé de dissolution de tissus cellulaires et de libération des cellules ou groupes de cellules contenus dans ces tissus par incubation du tissu cellulaire avec une protéase neutre recombinée de Clostridium histolyticum, codée par a) un ADN des nucléotides 1027-1965 de numéro d'identification de séquence 3 ou un ADN complémentaire, b) des acides nucléiques qui s'hybrident à un ADN des nucléotides 1027-1965 de numéro d'identification de séquence 3, c) des acides nucléiques, qui s'hybrideraient sans dégénérescence du code génétique à un des acides nucléiques mentionnés en a) ou b). Cette protéase est le produit d'une expression procaryote ou eucaryote d'un acide nucléique exogène. L'incubation est poursuivie jusqu'à libération de la quantité désirée de cellules ou groupes de cellules et jusqu'à ce que les cellules ou groupes de cellules se séparent des fractions de tissus cellulaires.

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