Modification of Bacterial Artificial Chromosomes (BACs) and Preparation of Intact BACDNA for Generation of Transgenic Mice
Open Access
- 1 April 2005
- journal article
- unit
- Published by Wiley in Current Protocols in Neuroscience
- Vol. 31 (1) , 5.21.1-5.21.13
- https://doi.org/10.1002/0471142301.ns0521s31
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.Keywords
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