SpringerProtocols: Abstract: 10. Copy-Control pBAC/oriV Vectors for Genomic Cloning

10. Copy-Control pBAC/oriV Vectors for Genomic Cloning

By: Jadwiga Wild³, Waclaw Szybalski³

Abstract

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The use of the improved BAC system for cloning genomic DNA and library constructions is described. This system retains all the advantages of the original BACs but, in addition, permits, on command, amplification of the BAC plasmids and cloned DNA. This system consists of (1) plasmid pBAC/oriV containing an additional replication origin, oriV, and (2) a host carrying the up-mutants of the trfA replicator gene expressed from the L-arabinose-inducible Para promoter. The pBAC/oriV clones are always maintained in the single-copy state, but if more DNA is required, they could be amplified up to 100-fold, depending on the size of the cloned insert.

Affiliation(s): (3) McArdle Laboratory for Cancer Research, University of Wisconsin Medical School, Madison, WI

Book Title: Recombinant Gene Expression: Reviews and Protocols

Series: Methods in Molecular Biology | Volume: 267 | Pub. Date: Jul-20-2004 | Page Range: 145-154 | DOI: 10.1385/1-59259-774-2:145

Subject: Genetics/Genomics

Key Words: BAC libraries - amplifiable copy number - F plasmid oriS/RepE replicon - oriV/TrfA replication

References

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