MGMT-based method for obtaining high yields of recombinant protein expression
AssigneeInstitut Pasteur
InventorsPhilippe Despres, Sylvie Paulous, Elodie Crublet
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Abstract
The present invention relates to a novel enhancer of protein production in host cells. It discloses a vector for expressing recombinant proteins in these cells, comprising a nucleotide sequence encoding a) a secretion peptidic signal, b) a 6-methylguanine-DNA-methyltransferase enzyme (MGMT, EC 2.1.1.63), a mutant or a catalytic domain thereof, and c) a recombinant protein. Said MGMT enzyme is preferably the so-called SNAP protein.
Claims17
A vector for expressing recombinant proteins in isolated host cells, comprising a nucleotide sequence encoding in a single open reading frame, from 5′ to 3′:
- a) a peptidic secretion signal which is functional in said isolated host cells,
- b) a 6-methylguanine-DNA-methyltransferase enzyme (MGMT) or mutant thereof, wherein said MGMT enzyme or MGMT mutant is the protein of SEQ ID NO:2, or a homologous sequence thereof that is at least 80% identical to SEQ ID NO:2, and
- c) a recombinant protein.
A method of enhancing expression of recombinant protein(s) comprising co-expressing said protein(s) as fusion protein(s) from a vector encoding from 5′ to 3′:
- a peptidic secretion signal,
- a 6-methylguanine-DNA-methyltransferase enzyme (MGMT) or mutant thereof, wherein said MGMT enzyme or MGMT mutant is the protein of SEQ ID NO:2, or a homologous sequence thereof that is at least 80% identical to SEQ ID NO:2, and
- the recombinant protein(s).
Dependencies are inferred from the claim text.