Skip to main content
Log in

Construction of a Fosmid genomic library of Streptomyces roseoflavus Men-myco-93-63

  • Research Article
  • Published:
Frontiers of Agriculture in China

Abstract

To clone the antibiotic biosynthesis gene cluster of Streptomyces roseoflavus Men-myco-93-63, we constructed a Fosmid genomic library. The genomic DNA of the strain Men-myco-93-63 was isolated by the modified CTAB procedure, and the size of most genomic DNA fragments was larger than 150 kb. Then, a Fosmid genomic library containing more than 6000 clones was constructed. The average size of the inserted DNA in recombinant plasmids was 38.1 kb, and the probability of harboring any gene in the genome of the strain Men-myco-93-63 was 99.99%. The library coverage was at least a 10-fold genome equivalent. Therefore, the constructed Fosmid library meets the requirements as a standard genomic library

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Cantor C R, Smith C L, Mathew M K (1988). Pulsed-field gel electrophoresis of very large DNA molecules. Annu Rev Biophys Biophys Chem, 17(1): 287–304

    Article  PubMed  CAS  Google Scholar 

  • Di D P (2006). The extraction of the antibiotic and related genes cloning from Streptomyces roseoflavus Men-myco-93-63. Dissertation for the Doctoral Degree, Baoding: Agricultural University of Hebei, 31–46 (in Chinese)

    Google Scholar 

  • Gravius B, Cullum J, Hranueli D (1994). High G + C-content DNA markers for pulsed-field gel electrophoresis. Biotechniques, 16(1): 52

    PubMed  CAS  Google Scholar 

  • Hopwood D A, Bibb M J, Chater K F, Kieser T, Bruton C J, Kieser H M, Lydiate D J, Smith C P, Ward J M, Schrempf H (1985). Genetic Manipulation of Streptomyces: A Laboratory Manual. Norwich: the John Innes Foundation, 156–157

  • Hopwood D A, Kieser T (1990). The Streptomyces genome. In: Drlica K, Riley M, eds. The Bacterial Chromosome. Washington DC: ASM Press, 147–162

    Google Scholar 

  • Hradecna Z, Wild J, Szybalski W (1998). Conditionally amplifiable inserts in pBAC vectors. Microb Comp Genom, 3: 58

    Google Scholar 

  • Liu D Q (1992). Biological control of Streptomyces scabies and other plant pathogens. Dissertation for the Doctoral Degree, USA: the University of Minnesota, 8–20

    Google Scholar 

  • Liu L Q (2006). The study on the chitinase gene and the antibiotic biosynthesis gene cluster of Streptomyces roseoflavus. Dissertation for the Doctoral Degree, Baoding: Agricultural University of Hebei, 48–77 (in Chinese)

    Google Scholar 

  • MacNeil T, Gibbons P H (1986). Characterization of the Streptomyces plasmid pVE1. Plasmid, 16(3): 182–194

    Article  PubMed  CAS  Google Scholar 

  • Maniloff J (1989). Anomalous values of Mycoplasma genomes sizes determined by pulse-field gel electrophoresis. Nucleic Acids Res, 17(3): 1268

    Article  PubMed  CAS  Google Scholar 

  • Meng Q F, Yang W X, Zhang Q L, Liu D Q (2004). Field evaluation of antagonistic Streptomyces Men-myco-93-63 preparation in biocontrol of cotton Verticillium Wilt. In: Proceedings of the 15th International Plant Protection Congress. Beijing: Foreign Languages Press, 153

    Google Scholar 

  • Murray M G, Thompson W F (1980). Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res, 8(19): 4321–4326

    Article  PubMed  CAS  Google Scholar 

  • Pospiech A, Neumann B (1995). A versatile quick-prep of genomic DNA from gram-positive bacteria. Trends Genet, 11(6): 217–218

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Fritsch E F, Maniatis T (1989). Molecular Cloning: A Laboratory Manual. 2nd ed. New York: CSH Laboratory Press, 457

    Google Scholar 

  • Smith C L, Matsumoto T, Niwa O, Klco S, Fan J B, Yanagida M, Cantor C R (1987). An electrophoretic karyotype for Schizosaccharomyces pombe by pulsed field gel electrophoresis. Nucleic Acids Res, 15(11): 4481–4489

    Article  PubMed  CAS  Google Scholar 

  • Sun W M (2009). Study on the antibiotic biosynthetic gene related linear plasmid of Streptomyces roseoflavus Men-myco-93-63. Dissertation for the Masteral Degree, Baoding: Agricultural University of Hebei, 3–4 (in Chinese)

    Google Scholar 

  • Sun W M, Guo W, Liu D Q (2008). Efficient detection and analysis of linear plasmid in Streptomyces Men-myco-93-63. Acta Microbiol Sin, 48(12): 1671–1674 (in Chinese)

    CAS  Google Scholar 

  • Tobias K, Mervyn J B, Mark J B, Keith F C, David A H (2000). Practional Streptomyces Genetics. Norwich: the John Innes Foundation, 170–171

    Google Scholar 

  • Wild J, Hradecna Z, Szybalski W (2001). Single-copy/high-copy (SC/HC) pBAC/oriV novel vectors for genomics and gene expression. Plasmid, 45: 142–143

    Google Scholar 

  • Wild J, Hradecna Z, Szybalski W (2002). Conditionally amplifiable BACs: switching from single-copy to high-copy vectors and genomic clones. Genome Res, 12(9): 1434–1444

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Wei Guo or Daqun Liu.

About this article

Cite this article

Sun, W., Guo, W., Liu, D. et al. Construction of a Fosmid genomic library of Streptomyces roseoflavus Men-myco-93-63. Front. Agric. China 5, 576–580 (2011). https://doi.org/10.1007/s11703-011-1119-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11703-011-1119-x

Keywords

Navigation