Expression of recombinant mature lysostaphin

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

IPC codes

C12N 9/48 (2006.01) C07H 21/04 (2006.01) C07K 14/00 (2006.01) C12N 1/00 (2006.01) C12N 1/20 (2006.01) C12N 9/52 (2006.01) A61K 38/00 (2006.01)

Type

Patent

Patent number

CA 2388003

Description

A portion of the lysostaphin gene of Staphylococcus simulans has been cloned and overexpressed in the cytoplasm of E.coli to yield mature lysostaphin, in the absence of preprolysostaphin and prolysostaphin, under the transcriptional control of an IPTG-inducible promoter and a ribosome binding site. IPTG induction of the transformed host cells produces intracellular, soluble, mature lysostaphin (27 kDa), in the complete absence of preprolysostaphin and prolysostaphin. The mature lysostaphin so formed does not require post- translational modification. The mature lysostaphin so formed can be used to treat and prevent staphylococcal infections.

L'invention concerne une partie du gène de la lysostaphine deStaphylococcus simulans qui a été clônée et surexprimée dans le cytoplasme d'E.Coli pour donner de la lysostaphine mûre, en l'absence de prépolysostaphine et de prolysostaphine, sous le contrôle transcriptionnel d'un promoteur inductible d'IPTG et d'un site de liaison de ribosome. L'induction d'IPTG des cellules hôtes transformées produit de la lysostaphine mûre, soluble et intracellulaire (27kDa) en l'absence totale de préprolysostaphine et de prolysostaphine. La lysostaphine mûre ainsi formée ne nécessite pas de modification post-translationnelle. La lysostaphine mûre ainsi formée permet de traiter et prévenir les infections par staphylocoques.

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