Unit

UNIT 23.11 Modification and Production of BAC Transgenes

  1. Yongsu Jeong,
  2. Douglas J. Epstein

Published Online: 1 AUG 2005

DOI: 10.1002/0471142727.mb2311s71

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Jeong, Y. and Epstein, D. J. 2005. Modification and Production of BAC Transgenes. Current Protocols in Molecular Biology. 71:23.11:23.11.1–23.11.15.

Author Information

  1. University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania

Publication History

  1. Published Online: 1 AUG 2005
  2. Published Print: JUL 2005

Abstract

Bacterial artificial chromosomes (BACs) are the vectors of choice for the construction of genomic DNA libraries and, as such, have proven instrumental in the generation of large-scale physical maps; positional cloning projects; and the sequencing of human, mouse, and a plethora of other genomes. A number of methods have recently been developed to modify BAC DNA (e.g., insertion, deletion, substitution), making BACs even more useful for functional genomic research. This unit describes two protocols for BAC modification in E. coli, one that allows for specific changes at a given DNA sequence and another that is more suited for rapid and nonspecific integration of foreign DNA (such as a reporter cassette) into a BAC insert. In addition, a simple and reliable method for preparing BAC DNA for pronuclear microinjection is also provided.

Keywords:

  • Bacterial artificial chromosomes (BACs);
  • Homologous recombination;
  • Transposon;
  • Recombineering;
  • Transgenic mouse