System for efficient isolation of genes using probes
AssigneeThe Board of Trustees of the Leland Stanford Jr. University
InventorsRonald W. Davis, Richard A. Young
Life of this patent
Estimating the term…
Abstract
Systems are provided for the dual purpose of cloning genes using antibodies as probes and isolating unknown proteins encoded by cloned DNA. The method employs a vector derived from phage which is used in combination with a high frequency lysogenic host. The vector is further characterized by having an inducible promoter regulating expression of a gene into which the foreign DNA may be introduced to produce a fused protein, and controlled induction of the prophage with rapid increase in copy number and high level transcription of foreign DNA. The technique is exemplified with a specific lambda phage construct in conjunction with the .beta.-galactosidase structural gene lacZ.
Claims2
.lambda.gt11.
A method for screening a genomic cDNA library which comprises:
- inserting foreign cDNA sequences into .lambda.gt11 phage vectors at a unique site in the .beta.-galactosidase gene of said .lambda.gt11 wherein expression results in production of fused polypeptides;
- transforming a high-frequency lysogenic bacterial host with said vectors containing said cDNA sequences;
- growing said host to high density and inducing said phage to high copy number with concomitant expression of said fused polypeptides;
- and
- isolating a fused expression product of said .beta.-galactosidase gene and cDNA.
Dependencies are inferred from the claim text.