| US 7,531,325 B2 | ||
| Method for cleavage of fusion proteins | ||
| Gijs Van Rooijen, Calgary (Canada); Joenel Alcantara, Calgary (Canada); and Maurice M. Moloney, Calgary (Canada) | ||
| Assigned to SemBioSys Genetics Inc., Calgary, Alberta (Canada) | ||
| Filed on Dec. 19, 2002, as Appl. No. 10/322,746. | ||
| Application 10/322746 is a continuation in part of application No. 09/402488, previously published as PCT/CA98/00398, filed on Apr. 23, 1998. | ||
| Claims priority of provisional application 60/044254, filed on Apr. 25, 1997. | ||
| Prior Publication US 2003/0166162 A1, Sep. 04, 2003 | ||
| Int. Cl. C12P 21/04 (2006.01); C12P 21/06 (2006.01); C12P 21/00 (2006.01) | ||
| U.S. Cl. 435—69.7 [435/69.1; 800/4; 800/7] | 18 Claims |
| 1. A method for the preparation of a recombinant polypeptide, comprising
a) transforming a non-human host cell an expression vector comprising:
(1) a nucleic acid sequence capable of regulating transcription in a host cell, operatively linked to
(2) a chimeric nucleic acid sequence encoding a fusion protein, wherein said chimeric nucleic acid sequence comprises (a)
a nucleic acid sequence encoding a pro-peptide sequence comprising SEQ ID NO:10, linked in reading frame to (b) a nucleic
acid sequence that is heterologous to the pro-peptide and that encodes the recombinant polypeptide, wherein the heterologous
nucleic acid sequence is located immediately downstream of the nucleic acid sequence encoding the pro-peptide; operatively
linked to
(3) a nucleic acid sequence encoding a termination region functional in said host cell,
b) growing the host cell to produce said fusion protein;
c) obtaining said fusion protein from said non-human host cell, and
d) contacting said fusion protein with a mature form of an autocatalytically maturing aspartic protease that cleaves the pro-peptide,
whereby said pro-peptide is cleaved from said fusion protein to release said recombinant polypeptide.
|