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Generation of BAC Transgenic Mice

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Bacterial Artificial Chromosomes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 256))

Abstract

Transgenic mice provide an important experimental system for studying the effects of gene overexpression and for analyzing the expression of well-characterized “single” genes that contribute to a particular trait. However, single-gene expression in transgenic (Tg) mice is not appropriate for the functional analysis of the many genes that may reside in a typical genetically mapped locus of the order of 1 cM. Bacterial artificial chromosome (BAC) vectors, which can accommodate large genomic intervals of up to 300 kb are appropriate for such studies (1). BACs of this size may contain several contiguous genes along with their cis-acting regulatory elements. As such, BACs are valuable tools in complementation or “rescue” experiments for identifying genes contributing to a genetically mapped quantitative trait of interest.

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References

  1. Shizuya, H, Birren, B., Kim, U. J., et al. (1992) Cloning and stable maintenance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an F-factor-based vector. Proc. Natl. Acad. Sci. USA 89, 8794–8797.

    Article  PubMed  CAS  Google Scholar 

  2. Nielsen, L. B., McCormick, S. P., Pierotti, V., et al. (1997) Human apolipoprotein B transgenic mice generated with 207-and 145-kilobase pair bacterial artificial chromosomes. Evidence that a distant 5′-element confers appropriate transgene expression in the intestine. J. Biol. Chem. 272, 29,752–29,758.

    Article  PubMed  CAS  Google Scholar 

  3. Yang, X. W., Model, P., and Heintz, N. (1997) Homologous recombination based modification in Escherichia coli and germline transmission in transgenic mice of a bacterial artificial chromosome. Nat. Biotechnol. 15, 859–865.

    Article  PubMed  CAS  Google Scholar 

  4. Nefedov, M., Williamson, R., and Ioannou, P. A. (2000) Insertion of diseasecausing mutations in BACs by homologous recombination in Escherichia coli. Nucl. Acids Res. (Online). 28, E79.

    Article  PubMed  CAS  Google Scholar 

  5. Lalioti, M., and Heath, J. (2001) A new method for generating point mutations in bacterial artificial chromosomes by homologous recombination in Escherichia coli. Nucl. Acids Res. (Online). 29, E14.

    Article  PubMed  CAS  Google Scholar 

  6. Lamb, B. T., Sisodia, S. S., Lawler, A. M., et al. (1993) Introduction and expression of the 400 kilobase amyloid precursor protein gene in transgenic mice [corrected]. Nat. Genet. 5, 22–30.

    Article  PubMed  CAS  Google Scholar 

  7. Antoch, M. P., Song, E. J., Chang, A. M., et al. (1997) Functional identification of the mouse circadian Clock gene by transgenic BAC rescue. Cell 89, 655–667.

    Article  PubMed  CAS  Google Scholar 

  8. King, D. P., Zhao, Y., Sangoram, A. M., et al. (1997) Positional cloning of the mouse circadian clock gene. Cell 89, 641–653.

    Article  PubMed  CAS  Google Scholar 

  9. Probst, F. J., Fridell, R. A., Raphael, Y., et al. (1998) Correction of deafness in shaker-2 mice by an unconventional myosin in a BAC transgene. Science 280, 1444–1447.

    Article  PubMed  CAS  Google Scholar 

  10. Means, G. D., Boyd, Y, Willis, C. R., and Derry, J. M. (2001) Transgenic rescue of the tattered phenotype by using a BAC encoding Ebp. Mamm. Genome. 12, 323–325.

    Article  PubMed  CAS  Google Scholar 

  11. Hogan, B., Beddington, R., Costantini, F., and Lacy, E. (1994) Manipulating the Mouse Embryo, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  12. Laird, P. W., Zijderveld, A., Linders, K., Rudnicki, M. A., Jaenisch, R., and Berns, A. (1991) Simplified mammalian DNA isolation procedure. Nucl. Acids Res. 19, 4293.

    Article  PubMed  CAS  Google Scholar 

  13. Chrast, R., Scott, H. S., and Antonarakis, S. E. (1999) Linearization and purification of BAC DNA for the development of transgenic mice. Transgen. Res. 8, 147–150.

    Article  CAS  Google Scholar 

  14. Boulukos, K. E. and Pognonec, P. (2001) MICE, a program to track and monitor animals in animal facilities. BMC Genet. 2, 4.

    Article  PubMed  CAS  Google Scholar 

  15. McKie, M. A. and Webb, S. (1999) LAMS-A laboratory animal management system. Mamm. Genome. 10, 349–351.

    Article  PubMed  CAS  Google Scholar 

  16. Huxley, C. (1998) Exploring gene function: use of yeast artificial chromosome transgenesis. Methods 14, 199–210.

    Article  PubMed  CAS  Google Scholar 

  17. Mullins, L. J., Kotelevtseva, N, Boyd, A. C, and Mullins, J. J. (1997) Efficient Cre-lox linearisation of BACs: applications to physical mapping and generation of transgenic animals. Nucl. Acids Res. 25, 2539–2540.

    Article  PubMed  CAS  Google Scholar 

  18. Jakobovits, A., Moore, A. L., Green, L. L., et al. (1993) Germ-line transmission and expression of a human-derived yeast artificial chromosome. Nature 362, 255–258.

    Article  PubMed  CAS  Google Scholar 

  19. Kaufman, R. M., Pham, C. T., and Ley, T. J. (1999) Transgenic analysis of a 100-kb human beta-globin cluster-containing DNA fragment propagated as a bacterial artificial chromosome. Blood 94, 3178–3184.

    PubMed  CAS  Google Scholar 

  20. Peterson, K. R., Navas, P. A., Li, Q., and Stamatoyannopoulos, G. (1998) LCR-dependent gene expression in beta-globin YAC transgenics: detailed structural studies validate functional analysis even in the presence of fragmented YACs. Human Molec. Genet. 7, 2079–2088.

    Article  CAS  Google Scholar 

  21. Don, R. H., Cox, P. T., Wainwright, B. J., Baker, K., and Mattick, J. S. (1991) ‘Touchdown’ PCR to circumvent spurious priming during gene amplification. Nucl. Acids Res. 19, 4008.

    Article  PubMed  CAS  Google Scholar 

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© 2004 Humana Press Inc., Totowa, NJ

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Marshall, V.M., Allison, J., Templeton, T., Foote, S.J. (2004). Generation of BAC Transgenic Mice. In: Zhao, S., Stodolsky, M. (eds) Bacterial Artificial Chromosomes. Methods in Molecular Biology, vol 256. Humana Press. https://doi.org/10.1385/1-59259-753-X:159

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  • DOI: https://doi.org/10.1385/1-59259-753-X:159

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-989-6

  • Online ISBN: 978-1-59259-753-6

  • eBook Packages: Springer Protocols

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