Cleavage of caulobacter produced recombinant fusion proteins

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/62 (2006.01) C07K 14/00 (2006.01) C07K 14/08 (2006.01) C07K 14/21 (2006.01) C07K 14/245 (2006.01) C12N 15/74 (2006.01)

Patent

CA 2334080

This invention provides a method for cleaving target proteins from <u>Caulobacter</u> S-layer protein under mild acid conditions. A fusion protein secreted by <u>Caulobacter</u> which includes a target protein and a <u>Caulobacter</u> S-layer secretion signal may be cleaved at an aspartate- proline dipeptide without solubilizing the fusion protein. This method may be carried out while the fusion protein is in an insoluble aggregate which facilitates recovery of the protein. This invention also provides a method of preparing a DNA construct for expression of the fusion protein and a method of preparing the fusion protein.

La présente invention concerne un procédé de clivage de protéines cibles à partir d'une protéine de couche superficielle de ?u?Colobacter?/u? dans des conditions acides douces. Une protéine hybride sécrétée par ?u?Colobacter?/u? et comprenant une protéine cible et un signal de sécrétion de couche superficielle de ?u?Colobacter?/u? peut subir un clivage au niveau d'un dipeptide aspartate-proline sans solubiliser la protéine hybride. Ce procédé peut être réalisé alors que la protéine hybride est un agrégat insoluble, facilitant ainsi la récupération de la protéine. Par ailleurs, cette invention concerne un procédé de préparation d'une structure d'ADN pour l'expression de la protéine hybride et un procédé de préparation de la protéine hybride.

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