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

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