Current Microbiology

, Volume 63, Issue 1, pp 68–74

Streptomyces autolyticus JX-47 Large-Insert Bacterial Artificial Chromosome Library Construction and Identification of Clones Covering Geldanamycin Biosynthesis Gene Cluster

  • Shikun Dai
  • Yongchang Ouyang
  • Guanghua Wang
  • Xiang Li
Article

DOI: 10.1007/s00284-011-9940-2

Cite this article as:
Dai, S., Ouyang, Y., Wang, G. et al. Curr Microbiol (2011) 63: 68. doi:10.1007/s00284-011-9940-2

Abstract

Geldanamycin belongs to benzoquinone ansamycin antibiotic and has potent antitumor activities. In this study, a bacterial artificial chromosome (BAC) library with an average insert size of up to 150 kb was constructed from genomic DNA of Streptomyces autolyticus JX-47. A genetic-screening strategy was established using BAC end-sequencing and three pairs of primers designed to target the remote regions, gdmA1, gdmA3 and gdmRI, of the geldanamycin gene cluster. Three clones covering geldanamycin biosynthesis gene cluster were obtained, which together spanned a 250-kb genomic region, and a 150227-bp insert in the clone p4E9 was sequenced. Comparison with the reported geldanamycin gene cluster sequences from S. hygroscopicus revealed that it had the same gene arrangement and high gene homology in the polyketide synthase (PKS) region and its downstream with 84–100% DNA identity and 81–100% amino acid (AA) identity. Its DNA homology with the whole gene cluster sequence from S. hygroscopicus strain 17997 reached 99% identity. However, upstream of the PKS region exhibited great diversity, where only ORF16 was conserved, and the other genes including gdmL and gdmX were displaced.

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Shikun Dai
    • 1
    • 2
  • Yongchang Ouyang
    • 3
  • Guanghua Wang
    • 1
  • Xiang Li
    • 1
  1. 1.Key Laboratory of Marine Bio-Resources Sustainable Utilization, Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of OceanologyChinese Academy of SciencesGuangzhouPeople’s Republic of China
  2. 2.Graduate University of Chinese Academy of SciencesBeijingPeople’s Republic of China
  3. 3.Guangzhou Medical CollegeGuangzhouPeople’s Republic of China

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