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The construction and characteristics of a BAC library for Cucumis sativus L. ‘B10’


Cloning using bacterial artificial chromosomes (BACs) can yield high quality genomic libraries, which are used for the physical mapping, identification and isolation of genes, and for gene sequencing. A BAC genomic library was constructed from high molecular weight DNA (HMW DNA) obtained from nuclei of the cucumber (Cucumis sativus L. cv. Borszczagowski; B10 line). The DNA was digested with the HindIII restriction enzyme and ligated into the pCC1BAC vector. The library consists of 34,560 BAC clones with an average insert size of 135 kb, and 12.7x genome coverage. Screening the library for chloroplast and mitochondrial DNA content indicated an exceptionally low 0.26% contamination with chloroplast DNA and 0.3% with mitochondrial DNA.



bacterial artificial chromosome


albumin from bovine serum


4′-6′-Diamidino-2-phenylindole dihydrochloride


high molecular weight DNA


low melting agarose


polymerase chain reaction


pulsed field gel electrophoresis




sequence-characterised amplified regions


  1. Staub, J.E. and Meglic, V. Molecular genetic markers and their legal relevance for cultigen discrimination: A case study in cucumber. Hort. Technol. 3 (1993) 291–300.

    Google Scholar 

  2. Arumuganathan, K. and Earle, E.D. Nuclear DNA content of some important plant species. Plant Mol. Biol. Rep. 9 (1991) 208–219.

    CAS  Google Scholar 

  3. Malepszy, S. and Niemirowicz-Szczytt, K. Sex determination in cucumber (Cucumis sativus L.) as a model system for molecular biology. Plant Sci. 80 (1991) 39–47.

    Article  Google Scholar 

  4. Nam, Y.W., Lee, J.R., Song, K.H., Lee, M.K., Robbins, M.D., Chung, S.M., Staub, J.E. and Zhang, H.B. Construction of two BAC libraries from cucumber (Cucumis sativus L.) and identification of clones linked to yield component quantitative trait loci. Theor. Appl. Genet. 111 (2005) 150–160.

    PubMed  Article  CAS  Google Scholar 

  5. Shizuya, H., Birren, B., Kim, U.J., Mancino, V., Slepak, T., Tachiiri, Y. and Simon, M. 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 (1992) 8794–8797.

    PubMed  Article  CAS  Google Scholar 

  6. Shultz, J., LaMontagne, C.H., Zobrist, K., Meksem K., Wu, C.H., Tao, Q., Nimmakayala, P., Santos, F., Zhang, H. and Lightfoot, D.A. Integrated physical mapping of the soybean genome: A tool for rapid identification of economically important genes. Soyb. Genet. Newsl. Vol. 27 (2000) 1–7.

    Google Scholar 

  7. Zhang, P., Li, W., Fellers, J., Friebe, B. and Gill, B.S. BAC-FISH in wheat identifies chromosome landmarks consisting of different types of transposable elements. Chromosoma 112 (2004) 288–299.

    PubMed  Article  CAS  Google Scholar 

  8. Peterson, D.G., Tomkins, J.P., Frisch, D.A., Wing, R.A. and Paterson, A.H. Construction of plant bacterial artificial chromosome (BAC) libraries: An illustrated guide. J. Agricult. Genomics 5 (2000) 1–100.

    Google Scholar 

  9. Sambrook, J., Fritsh, E.F. and Maniatis, T. Molecular cloning—A laboratory manual. (N Ford, C Nolan, M Ferguson Ed.) 2th edition, Cold Spring Harbor Lab. Pres (1989) New York.

    Google Scholar 

  10. Woo, S.S., Rastogi, V.K., Zhang, H.B., Paterson, A.H., Schertz, K.F. and Wing, R. Isolation of megabase-size DNA from sorghum sand applications for physical mapping and bacterial and yeast artificial chromosome library construction. Plant Mol. Biol. 13 (1995) 82–94.

    CAS  Google Scholar 

  11. Martin, G.B., Ganal, M.W. and Tanksley, S.D. Construction of a yeast artificial chromosome library of tomato and identification of cloned segments linked to two disease resistance loci. Mol. Gen. Genet. 233 (1992) 25–32.

    PubMed  Article  CAS  Google Scholar 

  12. Wing, R.A., Rastogi, V.K., Zhang, H.B., Paterson, A.H. and Tanksley, S.D. An improved method of plant megabase DNA isolation in agarose microbeads suitable for physical mapping and YAC cloning. Plant J. 4 (1993) 893–898.

    PubMed  Article  CAS  Google Scholar 

  13. Woo, S.S., Jiang, J., Gill, B.S., Paterson, A.H. and Wing, R.A. Construction and characterization of a bacterial artificial chromosome library of Sorghum bicolor. Nucleic Acid Res. 22 (1994) 4922–4931.

    PubMed  Article  CAS  Google Scholar 

  14. Peterson, D.G., Boehm, K.S. and Stack, S.M. Isolation of milligram quantities of DNA from tomato (Lycopersicon esculentum), a plant containing high levels of polyphenolic compounds. Plant Mol. Biol. 15 (1997) 148–153.

    CAS  Google Scholar 

  15. Peterson, D.G., Pearson, W.R. and Stack, S.M. Characterization of the tomato (Lycopersicon esculentum) genome using in vitro and in situ DNA reassociation. Genome 41 (1998) 346–356.

    Article  CAS  Google Scholar 

  16. Tomkins, J.P., Peterson, D.G., Yang, T.J., Main, D., Wilkins, T.A., Paterson, A.H. and Wing, R.A. Development of genomic resources for cotton (Gossypium hirsutum L.): BAC library construction, preliminary STC analysis, and identification of clones associated with fiber development. Mol. Breed. 8 (2001) 255–261.

    Article  CAS  Google Scholar 

  17. Watson, J.C. and Thompson, W.F. Purification and restriction endonuclease analysis of plant nuclear DNA. Meth. Enzymol. 118 (1986) 57–79.

    CAS  Google Scholar 

  18. Song, B.K., Nadarajah, K., Romanov, M.N. and Ratnam, W. Application of overgo-based hybridization method in screening oryza rufipogon bacterial artificial chromosome (BAC) library. Cell. Mol. Biol. Lett. Vol. 10 (2005) 425–437.

    PubMed  CAS  Google Scholar 

  19. Choi, S., Creelman, R.A., Mullet, J.E., Wing, R.A. Construction and characterization of a bacterial artificial chromosome library of Arabidopsis thaliana. Plant Mol. Biol. 13 (1995) 124–128.

    Google Scholar 

  20. Cenci, A., Chantret, N., Kong, X., Gu, Y., Anderson, O.D., Fahima, T., Distelfeld, A. and Dubcovsky, J. Construction and characterization of a half million clone BAC library of durum wheat (Triticum turgidum ssp. durum). Theor. Appl. Genet. 107 (2003) 931–939.

    PubMed  Article  CAS  Google Scholar 

  21. Yu, Y., Tomkins, J.P., Waugh, R., Frich, D.A., Kudrna, D., Kleinhofs, A., Brueggeman, R.S., Muehlbauer, G.J., Wise, R.P. and Wing, R.A. A bacterial artificial chromosome library for barley (Hordeum vulgare L.) and the identification of clones containing putative resistance genes. Theor. Appl. Genet. 101 (2000) 1093–1099.

    Article  CAS  Google Scholar 

  22. Burke, D.T. and Olsen, M.V. Preparation of clone libraries in yeast artificial chromosome vectors. Meth. Enzymol. 194 (1991) 251–270.

    PubMed  CAS  Article  Google Scholar 

  23. Anand, R., Viasante, A. and Tyler-Smith, C. Construction of yeast artificial chromosome libraries with large insert using fractionation by pulsed-field gel electrophoresis. Nucleic Acids Res. 17 (1989) 3425–3433.

    PubMed  Article  CAS  Google Scholar 

  24. Clarke, L. and Carbon, J. A colony bank containing synthetic CoIEI hybrid plasmids representative of the entire E. coli genes. Cell 9 (1976) 91–99.

    PubMed  Article  CAS  Google Scholar 

  25. Hamilton, C.M., Frary, A., Xu, Y., Tanksley, S.D. and Zhang, H.B. Construction of tomato genomic DNA library in a binary-BAC (BIBAC) vector. Plant J. 18 (1999) 223–229.

    Article  CAS  Google Scholar 

  26. Wu, C.C., Nimmakayala, P., Santos, F.A., Springman, R., Scheuring, C., Meksem, K., Lightfoot, D.A. and Zhang, H.B. Construction and characterization of a soybean bacterial artificial chromosome library and use of multiple complementary libraries for genome physical mapping. Theor. Appl. Genet. 109 (2004) 1041–1050.

    PubMed  Article  CAS  Google Scholar 

  27. Blanchard, J.L. and Schmidt, G.W. Pervasive migration of organellar DNA to the nucleus in plants. J. Mol. Evol. 4 (1995) 397–406.

    Article  Google Scholar 

  28. Przybecki, Z., Kowalczyk, M.E., Witkowicz, J., Filipecki, M. and Siedlecka, E. Polymorphom of sexually different cucumber (Cucumis sativus L.) NIL lines. Cell. Mol. Biol. Lett. 9 (2004) 919–933.

    PubMed  CAS  Google Scholar 

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

    PubMed  Article  CAS  Google Scholar 

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Correspondence to Wojciech Gutman.

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Gutman, W., Pawełkowicz, M., Woycicki, R. et al. The construction and characteristics of a BAC library for Cucumis sativus L. ‘B10’. Cell Mol Biol Lett 13, 74–91 (2008).

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Key words

  • Cucumis sativus L.
  • Bacterial artificial chromosome
  • High molecular weight DNA
  • Pulsed field gel electrophoresis