Unit

UNIT 5.21 Modification of Bacterial Artificial Chromosomes (BACs) and Preparation of Intact BAC DNA for Generation of Transgenic Mice

  1. Shiaoching Gong1,
  2. X. William Yang2

Published Online: 1 MAY 2005

DOI: 10.1002/0471142301.ns0521s31

Current Protocols in Neuroscience

Current Protocols in Neuroscience

How to Cite

Gong, S. and Yang, X. W. 2005. Modification of Bacterial Artificial Chromosomes (BACs) and Preparation of Intact BAC DNA for Generation of Transgenic Mice. Current Protocols in Neuroscience. 31:5.21:5.21.1–5.21.13.

Author Information

  1. 1

    Rockefeller University, New York, New York

  2. 2

    Department of Psychiatry and Biobehavioral Sciences, Center for Neurobehavioral Genetics, Neuropsychiatric Institute, David Geffen School of Medicine at UCLA, Los Angeles, California

Publication History

  1. Published Online: 1 MAY 2005
  2. Published Print: APR 2005

Abstract

BAC transgenesis is a powerful tool for the study of gene expression and gene function in the mouse in vivo. In this unit, detailed protocols are provided for modification (i.e., marker gene insertion, deletion, or point mutation) of BACs by homologous recombination in E. coli. This method utilizes a shuttle vector that allows transient expression of the E. coli RecA gene to support homologous recombination in the BAC host bacteria. In addition, two protocols are provided for purification of BAC DNA for microinjection to generate transgenic mice. Since BAC DNA is prone to degradation, which may introduce positional effects in transgenic mice, two methods are given for purification of intact BAC DNA for subsequent microinjection.