African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12488

Full Length Research Paper

Construction of intergeneric conjugal transfer for molecular genetic studies of Streptomyces mobaraensis producing transglutaminase

Jae-Young Park
  • Jae-Young Park
  • Department of Food Science and Biotechnology, Kyungnam University, Changwon 631-701, South Korea.
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Sun-Uk Choi*
  • Sun-Uk Choi*
  • Department of Food Science and Biotechnology, Kyungnam University, Changwon 631-701, South Korea.
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  •  Received: 25 December 2013
  •  Accepted: 14 March 2014
  •  Published: 26 March 2014

Abstract

To facilitate molecular studies of Streptomyces mobaraensis producing transglutaminase, an effective transformation method was established via intergeneric conjugal transfer using Escherichia coli ET12567 harboring the ØC31-derived integration vector, pSET152. The highest frequency was attained on ISP4 medium containing 20 mM MgCl2, using 2.5 × 108 E. coli as donor and spore treated with heat treatment at 35°C for 10 min as host. The attB integration site in the S. mobaraensis genome was detected as a single attB site within an open reading frame coding for a pirin homolog. Its sequence showed the highest degree of homology with S. aureofaciens.

 

Key words: Streptomyces mobaraensis, conjugal transfer, integration site, exoconjugant, transglutaminase.

Abbreviation

Abbreviations: ORF, Open reading frame; int, integrase gene; ATTPR, attP right; ATTPL, attP left; ISP, international streptomyces project medium; AS-1, amino acid soluble starch medium; MS, mannitol soya flour medium.

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This article is published under the terms of the Creative Commons Attribution License 4.0

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