Abstract
Native plasmid of Streptococcus thermophilus ST137, pER371 (2.7 kb) linearized at various unique restriction sites was individually subcloned into Escherichia coli plasmid pUC19 to generate the pUER-series recombinants. A selection cassette consisting of chloramphenicol- and erythromycin-resistance genes was spliced into each construct to generate the pMEU shuttle vectors. Electrotransformation of Streptococcus thermophilus with these vectors showed that a ca. 1.7 kb BstEII/ BanII fragment is essential for plasmid replication. A shuttle vector, pMEU149-1 (5.3 kb), was constructed using the minimally required fragment for replication. A chloramphenicol acetyltransferase ( cat) gene was successfully expressed in the ultimate S. thermophilus host by using pMEU149-1. Cloning vectors derived from pER371 should provide valuable alternative gene delivery vehicles for the genetic engineering of lactic acid bacteria.
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Solaiman, D.K., Somkuti, G.A. Characterization of pER371-based Streptococcus thermophilus-Escherichia coli shuttle vectors. Biotechnology Letters 19, 595–598 (1997). https://doi.org/10.1023/A:1018362025665
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DOI: https://doi.org/10.1023/A:1018362025665