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US 4,366,246 AGranted
Method for microbial polypeptide expression
AssigneeGenentech, Inc.
InventorArthur D. Riggs
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AssigneeGenentech, Inc.
InventorArthur D. Riggs
Estimating the term…
The Specification discloses: 1. Recombinant microbial cloning vehicles comprising heterologous DNA coding for the expression of mammalian hormone (e.g., somatostatin) and other polypeptides, including plasmids suited for the transformation of bacterial hosts. The latter incorporate a regulon homologous to the host in its untransformed state, in reading phase with the structural gene for the heterologous DNA; 2. Cloning vehicles coding for the microbial expression of a protein variously comprising (a) a polypeptide hapten and additional protein sufficient in size to confer immunogenicity on the product of expression, which may find use in raising antibodies to the hapten for assay use or in the manufacture of vaccines; and (b) a desired polypeptide product and additional protein from which the desired product may be cleaved; and 3. Methods of preparing synthetic structural genes coding for the expression of mammalian polypeptides in microbial cloning systems.
In the method of producing a specific polypeptide involving expression of a heterologous structural gene therefor in a recombinant microbial cloning vehicle, the method comprising the steps of growing microbial transformants comprising such structural gene-containing vehicles and expressing the amino acid sequence encoded by such gene, the improvement wherein the structural gene is in reading phase with a DNA sequence coding for a protein other than said polypeptide so that expression yields a precursor protein comprising both the amino acid sequence of the polypeptide and additional protein containing a selective cleavage site adjacent the amino acid sequence of said specific polypeptide.
In the method of producing a specific mammalian polypeptide hormone or intermediate therefor involving expression of a heterologous structural gene therefor in a recombinant bacterial plasmid wherein the structural gene is followed by one or more termination codons, the method comprising the steps of growing bacterial transformants comprising such structural gene-containing plasmids and expressing the amino acid sequence encoded by such gene, the improvement wherein the structural gene is preceded by and in reading phase with a DNA sequence coding for a protein other than said polypeptide so that expression yields a precursor protein comprising both the amino acid sequence of the polypeptide and additional protein containing a selective cleavage site adjacent said polypeptide's amino acid sequence, whereafter cleavage is effected in a system exogenous to the replicative environment of the plasmid.
Dependencies are inferred from the claim text.