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Construction of intergeneric conjugal transfer for molecular genetic studies of <i>Streptomyces mobaraensis</i> producing transglutaminase


Jae-Young Park
Sun-Uk Choi

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.

Keywords: Streptomyces mobaraensis, conjugal transfer, integration site, exoconjugant, transglutaminase

African Journal of Biotechnology, Vol 13(12), 1462-1466

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eISSN: 1684-5315