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US 2010/0136692 A1Published application
Plasmids and phages for homologous recombination and methods of use
AssigneeThe Govt. of the U.S.A. as represented by the Secretary of the Dept. of Health & Human Services
InventorsDonald L. Court, Simanti Datta, Nina Costantino
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Abstract
Lambda phages that can be used to introduce recombineering functions into host cells are disclosed. Also disclosed are plasmids that can be used to confer recombineering functions to a variety of strains of E. coli and to other bacteria, including Salmonella, Pseudomonas, Cyanobacteria, Spirochaetes. These plasmids and phages can be isolated in vitro and can be used to transform bacterial cells, such as gram negative bacteria.
Claims20
A lambda phage comprising,
- a repressor that binds a PL promoter;
- a promoter operably linked to a nucleic acid encoding a heterologous nucleic acid sequence;
- PL, and a nucleic acid encoding Beta operably linked to PL;
- a nucleic acid encoding P, a nucleic acid encoding O, a nucleic acid encoding Cro;
- and a phage origin of replication,
- wherein at least two of the nucleic acids encoding P, encoding O, and encoding Cro comprise an amber codon such that at least two of P, O, and Cro proteins are not produced when the lambda phage is introduced into a suppressor minus host cell;
- and
- and wherein the lambda phage does not comprise a bacterial origin of replication.
5 more elements
Dependencies are inferred from the claim text.