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Characterization of pER371-based Streptococcus thermophilus-Escherichia coli shuttle vectors



Characterization of pER371-based Streptococcus thermophilus-Escherichia coli shuttle vectors



Biotechnology letters 19(7): 595-598



Native plasmid of Streptococcus thermophilus ST 137, 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, pMEU14'-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 pMEU14'-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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Accession: 002775847

Download citation: RISBibTeXText

DOI: 10.1023/a:1018362025665


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