Expression systems for amidating enzyme
AssigneeUnigene Laboratories Inc.
InventorsArthur H. Bertelsen, Nozer M. Mehta, Gary Agide Beaudry, James P. Gilligan, Barry N. Jones
Life of this patent
Estimating the term…
Abstract
Alpha-amidating enzyme is produced by recombinant DNA techniques recoverable in high yields and at high purity. Both eukaryotic and prokaryotic expression vectors are provided having a transcriptional promoter followed downstream by a DNA sequence which encodes amidating enzyme. The vector selected is one capable of directing the expression of polypeptides in the host selected, and preferred hosts are transected with the described vectors.
Claims38
A host cell transformed or transfected with an expression vector comprising a first DNA sequence which hybridizes under stringent conditions with a natural DNA sequence shown in FIGS. 5A-5F.
A host cell transformed or transfected with an expression vector comprising a first DNA sequence which hybridizes under stringent conditions with a natural DNA sequence.
The host cell of claim 21 or 22, wherein said host cell is a chinese hamster ovary cell.
An expression vector for directing the expression of a polypeptide sequence of an alpha amidating enzyme in prokaryotic cells, said vector comprising a first DNA sequence having an amidating enzyme-coding region, said first DNA sequence being sufficiently homologous to a natural DNA sequence for encoding natural amidating enzyme to undergo hybridization with said natural sequence under stringent conditions, wherein said natural DNA sequence is shown in FIGS. 5A-5F.
An expression vector for directing the expression of a polypeptide sequence of an alpha amidating enzyme in prokaryotic cells, said vector comprising a first DNA sequence having an amidating enzyme-coding region, said first DNA sequence being sufficiently homologous to a natural DNA sequence for encoding natural amidating enzyme to undergo hybridization with said natural sequence under stringent conditions, wherein said natural DNA sequence is shown in FIGS. 6A-6F.
An expression vector for directing the expression of a polypeptide sequence of an alpha amidating enzyme in eukaryotic cells, said vector comprising a first DNA sequence having an amidating enzyme-coding region, said first sequence being sufficiently homologous to a natural DNA sequence for expressing natural amidating enzyme to undergo hybridization with said natural sequence under stringent conditions, and said first sequence including a stop codon 5' of a membrane spanning domain, wherein said natural DNA sequence is shown in FIGS. 5A-5F.
An expression vector for directing the expression of a polypeptide sequence of an alpha anidating enzyme in eukaryotic cells, said vector comprising a first DNA sequence having an anidating enzyme-coding region, said first sequence being sufficiently homologous to a natural DNA sequence for expressing natural amidating enzyme to undergo hybridization with said natural sequence under stringent conditions, and said first sequence including a stop codon 5' of a membrane spanning domain, wherein said natural DNA sequence is shown in FIGS. 6A-6F.
A host cell for expressing the polypeptide sequence of an alpha amdiating enzyme, said host cell transformed or transfected with an expression vector containing the DNA sequence shown in FIGS. 5A-5F.
A host cell for expressing the polypeptide sequence of an alpha amidating enzyme, said host cell transformed or transfected with an expression vector containing the DNA sequence shown in FIG. 6.
A method of manufacturing an amidated peptide comprising contacting a precursor of said peptide with a peptidylglyuine alpha-amidating monooxygenase expressed from a host cell transformed or transfected with an expression vector comprising a first,DNA sequence which hybridizes under stringent conditions with a natural DNA sequence that is shown in FIGS. 5A-5F or FIGS. 6A-6F.
Dependencies are inferred from the claim text.