Skip to main content

Construction of Small Genome BAC Library for Functional and Genomic Applications

  • Protocol
Bacterial Artificial Chromosomes

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 255))

  • 1419 Accesses

Abstract

The use of genetic approaches to study bacteria is limited by the following considerations: first, the bacteria must be easy to culture and nonpathogenic; and, second, a broad spectrum of genetic techniques for the particular bacteria being studied must be available. For other bacteria, such as the syphilis-causing spirochete, Treponema pallidum, genetic studies are highly limited because the bacterium cannot be continuously grown in the laboratory, because of its high virulence and a total lack of genetic tools to study this organism.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Brosch, R., Gordon, S. V., Billault, A., et al. (1998) Use of a Mycobacterium tuberculosis H37Rv bacterial artificial chromosome library for genome mapping, sequencing, and comparative genomics. Infect. Immun. 66, 2221–2229.

    PubMed  CAS  Google Scholar 

  2. Dewar, K., Sabbagh, L., Cardinal, G., Veilleux, F., Sanschagrin, F., Birren, B., and Levesque, R. C. (1998) Pseudomonas aeruginosa PAO1 bacterial artificial chromosomes: strategies for mapping, screening, and sequencing 100 kb loci of the 5.9 Mb genome. Microb. Comp. Genomics 3, 105–117.

    PubMed  CAS  Google Scholar 

  3. Tomkins, J. P., Miller-Smith, H., Sasinowski, M., et al. (1999) Physical map and gene survey of the Ochrobactrum anthropi genome using bacterial artificial chromosome contigs. Microb. Comp. Genomics 4, 203–217.

    PubMed  CAS  Google Scholar 

  4. Xu, Y., Murray, B. E., and Weinstock, G. M. (1998) A cluster of genes involved in polysaccharide biosynthesis from Enterococcus faecalis OG1RF. Infect. Immun. 66, 4313–4323.

    PubMed  CAS  Google Scholar 

  5. Rondon, M. R., Raffel, S. J., Goodman, R. M., and Handelsman, J. (1999) Toward functional genomics in bacteria: analysis of gene expression in Escherichia coli from a bacterial artificial chromosome library of Bacillus cereus. Proc. Natl. Acad. Sci. USA 96, 6451–6455.

    Article  PubMed  CAS  Google Scholar 

  6. Kim, U. J., Birren, B. W., Slepak, T., et al. (1996) Construction and characterization of a human bacterial artificial chromosome library. Genomics 34, 213–218.

    Article  PubMed  CAS  Google Scholar 

  7. Baseman, J. B. and Hayes, N. S. (1974) Protein synthesis by Treponema pallidum extracted from infected rabbit tissue. Infect. Immun. 10, 1350–1355.

    PubMed  CAS  Google Scholar 

  8. Hanff, P. A., Norris, S. J., Lovett, M. A., and Miller, J. N. (1984) Purification of Treponema pallidum, Nichols strain, by Percoll density gradient centrifugation. Sex. Transm. Dis. 11, 275–286.

    Article  PubMed  CAS  Google Scholar 

  9. Strong, S. J., Ohta, Y., Litman, G. W., and Amemiya, C. T. (1997) Marked improvement of PAC and BAC cloning is achieved using electroelution of pulsed-field gel-separated partial digests of genomic DNA. Nucleic Acids Res. 25, 3959–3961.

    Article  PubMed  CAS  Google Scholar 

  10. Fraser, C. M., Norris, S. J., Weinstock, G. M., et al. (1998) Complete genome sequence of Treponema pallidum, the syphilis spirochete. Science 281, 375–388.

    Article  PubMed  CAS  Google Scholar 

  11. Osoegawa, K., Woon, P. Y., Zhao, B., Frengen, E., Tateno, M., Catanese, J. J., and de Jong, P. J. (1998) An improved approach for construction of bacterial artificial chromosome libraries. Genomics 52, 1–8.

    Article  PubMed  CAS  Google Scholar 

  12. Zhu, H. and Dean, R. A. (1999) A novel method for increasing the transformation efficiency of Escherichia coli—application for bacterial artificial chromosome library construction. Nucleic Acids Res. 27, 910, 911.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2004 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Šmajs, D., Norris, S.J., Weinstock, G.M. (2004). Construction of Small Genome BAC Library for Functional and Genomic Applications. In: Zhao, S., Stodolsky, M. (eds) Bacterial Artificial Chromosomes. Methods in Molecular Biology™, vol 255. Humana Press. https://doi.org/10.1385/1-59259-752-1:047

Download citation

  • DOI: https://doi.org/10.1385/1-59259-752-1:047

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-988-9

  • Online ISBN: 978-1-59259-752-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics