Method of using bacteriophage lambda p.sub.1 promoter to produce a functional polypeptide in streptomyces
AssigneeEli Lilly and Company
InventorsStuart A. Kuhstoss, R. Nagaraja Rao
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Abstract
The invention relates to a method and cloning vehicle for the expression of a functional polypeptide in Streptomyces. A recombinant DNA cloning vehicle was genetically engineered to bring the expression of the neomycin phosphotransferase gene under the control of the Escherichia coli bacteriophage .lambda.p.sub.L promoter.
Claims17
A method of using bacteriophage lambda p.sub.L promoter to produce a functional polypeptide in Streptomyces, said method comprising:
- (a) transforming a Streptomyces host cell with a selectable and autonomously replicating recombinant DNA expression vector comprising:
- (1) the bacteriophage .lambda.p.sub.L promoter-operator transcriptional activating sequence;
- (2) a translational activating sequence;
- (3) a DNA sequence that codes for a functional polypeptide;
- and
- (b) culturing said transformed cell under conditions suitable for expression of said polypeptide,
- subject to the limitation that said expression vector sequentially contains said .lambda.p.sub.L promoter-operator transcriptional activating sequence, said translational activating sequence and said DNA sequence that codes for a functional polypeptide such that a translatable mRNA transcript encodes the functional polypeptide.
A plasmid selected from the group consisting of pKC309, pKC322, pKC283, pKC354, and pKC356.
A Streptomyces host cell transformed by a recombinant DNA expression vector capable of selection and autonomous replication in said Streptomyces host cell, said vector comprising:
- (a) the bacteriophage .lambda.P.sub.L promoter-operator transcription activating sequence;
- (b) a translational activating sequence;
- and
- (c) a DNA sequence that codes for a functional polypetide,
- subject to the limitation that said expression vector sequentially contains said .lambda.P.sub.L promoter-operator transcriptional activating sequence, said translational activating sequence, and said DNA sequence that codes for a functional polypeptide such that a translatable mRNA transcript encodes said functional polypeptide.
A selectable recombinant DNA expression vector which replicates in Streptomyces which comprises:
- (a) the bacteriophage .lambda.P.sub.L promoter-operator transcriptional activating sequence;
- (b) a translational activating sequence;
- and
- (c) a DNA sequence that codes for a functional polypeptide,
- subject to the limitation that in a Streptomyces host cell said expression vector sequentially contains said .lambda.P.sub.L promoter-operater transcriptional activating sequence, said translational activating sequence, and said DNA sequence that codes for a functional polypeptide such that a translatable mRNA transcript encodes said functional polypeptide.
Dependencies are inferred from the claim text.