+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Modification of bacterial artificial chromosomes (BACs) and preparation of intact BAC DNA for generation of transgenic mice

Modification of bacterial artificial chromosomes (BACs) and preparation of intact BAC DNA for generation of transgenic mice

Current Protocols in Neuroscience Chapter 5: Unit 5.21

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.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 054421818

Download citation: RISBibTeXText

PMID: 18428623

DOI: 10.1002/0471142301.ns0521s31

Related references

Efficient generation of transgenic mice with intact yeast artificial chromosomes by intracytoplasmic sperm injection. Biology of Reproduction 71(6): 1943-1947, 2004

Generation of Cre recombinase-expressing transgenic mice using bacterial artificial chromosomes. Methods in Molecular Biology 530: 325-342, 2009

Highly efficient modification of bacterial artificial chromosomes (BACs) using novel shuttle vectors containing the R6Kgamma origin of replication. Genome Research 12(12): 1992-1998, 2002

Generation of transgenic mice with yeast artificial chromosomes. Cold Spring Harbor Symposia on Quantitative Biology 58: 55-62, 1993

Framework for a physical map of the human 22q13 region using bacterial artificial chromosomes (BACs). Genomics 33(1): 9, 1996

Fluorescence in situ hybridization with Bacterial Artificial Chromosomes (BACs) to mitotic heterochromatin of Drosophila. Methods in Molecular Biology 659: 389-400, 2010

Direct sequencing of bacterial artificial chromosomes (BACs) and prokaryotic genomes by biotin-capture PCR. Journal of Biotechnology 60(1-2): 119-129, 1998

Generation of transgenic rats with YACs and BACs: preparation procedures and integrity of microinjected DNA. Experimental Animals 49(3): 229-233, 2000

Generation of bacterial artificial chromosome (BAC) transgenic mice. Methods in Molecular Biology 1194: 157-169, 2014

Pronuclear microinjection of purified artificial chromosomes for generation of transgenic mice: pick-and-inject technique. Methods in Molecular Biology 240: 227-242, 2004

Bacterial artificial chromosomes (BACs)-on-Beads™ as a diagnostic platform for the rapid aneuploidy screening of products of conception. Molecular Medicine Reports 8(2): 650-654, 2013

Construction and characterization of bacterial artificial chromosomes (BACs) containing herpes simplex virus full-length genomes. Methods in Molecular Biology 1144: 43-62, 2014

Generating transgenic mice from bacterial artificial chromosomes: transgenesis efficiency, integration and expression outcomes. Transgenic Research 18(5): 769-785, 2009

Generation of inner ear-specific knockout mice by modification of a Pax-2 bacterial artificial chromosome. Developmental Biology 259(2): 553, 2003

Human apolipoprotein B transgenic mice from 210-kb and 145-kb bacterial artificial chromosomes Apo-B gene expression in the intestine is controlled by a distant regulatory element. Circulation 94(8 Suppl. ): I631, 1996