Construction of Small Genome BAC Library for Functional and Genomic Applications

  • David Šmajs
  • Steven J. Norris
  • George M. Weinstock
Part of the Methods in Molecular Biology™ book series (MIMB, volume 255)


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.


Bacterial Artificial Chromosome Luria Bertani Bacterial Artificial Chromosome Clone Bacterial Artificial Chromosome Library White Coloni 
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  1. 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.PubMedGoogle Scholar
  2. 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.PubMedGoogle Scholar
  3. 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.PubMedGoogle Scholar
  4. 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.PubMedGoogle Scholar
  5. 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.PubMedCrossRefGoogle Scholar
  6. 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.PubMedCrossRefGoogle Scholar
  7. 7.
    Baseman, J. B. and Hayes, N. S. (1974) Protein synthesis by Treponema pallidum extracted from infected rabbit tissue. Infect. Immun. 10, 1350–1355.PubMedGoogle Scholar
  8. 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.PubMedCrossRefGoogle Scholar
  9. 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.PubMedCrossRefGoogle Scholar
  10. 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.PubMedCrossRefGoogle Scholar
  11. 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.PubMedCrossRefGoogle Scholar
  12. 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.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2004

Authors and Affiliations

  • David Šmajs
    • 1
  • Steven J. Norris
    • 2
  • George M. Weinstock
    • 3
  1. 1.Department of Microbiology and Molecular GeneticsUniversity of Texas-Houston Medical SchoolHouston
  2. 2.Department of Pathology and Laboratory MedicineUniversity of Texas-Houston Medical SchoolHouston
  3. 3.Human Genome Sequencing CenterBaylor College of MedicineHouston

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