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Development of a Plasmid Vector for Easy Selection of Strong Promoters

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Abstract.

A promoter vector pACPR33 for Escherichia coli based on the promotorless ampicillin-resistance gene from pBR322 has been constructed. The promoter of the ampicillin-resistance gene was deleted and replaced by a suitable multiple cloning site. Molecular cloning of promoters into the polylinker resulted in activation of the ampicillin resistance in E. coli. The plasmid contains a functional origin of DNA replication and a tetracycline resistance gene for E. coli, and a chloramphenicol resistance gene for S. aureus. The vector permitted direct detection of promoter activity, especially strong promoters, by easy iodometric determination of β-lactamase activity in liquid or solid media.

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Authors and Affiliations

  1. Laboratorio BioSidus S.A., (1254) Constitución 4234, Buenos Aires, Argentina , , , , , , AR

    Silvia A. Piñeiro

  2. Departamento de Microbiología, Parasitología e Inmunología, Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155 P-12, 1121 Buenos Aires, Argentina , , , , , , AR

    Daniel O. Sordelli & Daniela Centrón

Authors
  1. Silvia A. Piñeiro
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  2. Daniel O. Sordelli
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  3. Daniela Centrón
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Received: 26 July 1999 / Accepted: 22 November 1999

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Piñeiro, S., Sordelli, D. & Centrón, D. Development of a Plasmid Vector for Easy Selection of Strong Promoters. Curr Microbiol 40, 302–305 (2000). https://doi.org/10.1007/s002849910060

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  • DOI: https://doi.org/10.1007/s002849910060

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