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US 7,608,448 B2Granted
Method and cloning vectors utilizing intergeneric conjugation for manipulation of actinomycetes biosynthesis genes
AssigneeWyeth
InventorNathan Magarvey
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Abstract
The present invention relates to a vector comprising sequences that permit direct transfer of the vector from one prokaryotic cell to another, such as by intergeneric conjugation. The invention also relates to methods of making and using the vector.
Claims18
An intergeneric shuttle vector comprising an expression control sequence;
- a multiple cloning site into which a nucleic acid encoding a polypeptide can be inserted that is positioned such that when the nucleic acid is inserted into the multiple cloning site the nucleic acid is operatively associated with the expression control sequence;
- an E. coli origin of replication;
- an actinomycetes origin of replication from pJV1;
- a cos cosmid cloning site;
- and an origin of transfer (OriT) that directs transfer of the vector from an E. coli cell to an actinomycetes cell during conjugation, and wherein the shuttle vector has a vector map according to pNWA200.
2 more elements
An intergeneric shuttle vector comprising an expression control sequence;
- a multiple cloning site into which a nucleic acid encoding a polypeptide can be inserted that is positioned such that when the nucleic acid is inserted into the multiple cloning site said nucleic acid is operatively associated with the expression control sequence;
- an E. coli origin of replication;
- an actinomycetes origin of replication from pJV1;
- a cos cosmid cloning site;
- and an origin of transfer (OriT) that directs transfer of the shuttle vector from an E. coli cell to an actinomycetes cell during conjugation, and wherein the shuttle vector has a vector map according to pNWA115.
2 more elements
Dependencies are inferred from the claim text.