Development of a Plasmid Vector for Easy Selection of Strong Promoters |
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| Authors: | Silvia A Piñeiro Daniel O Sordelli Daniela Centrón |
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| Institution: | (1) Laboratorio BioSidus S.A., (1254) Constitución 4234, Buenos Aires, Argentina , AR;(2) Departamento de Microbiología, Parasitología e Inmunología, Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155 P-12, 1121 Buenos Aires, Argentina , AR |
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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.
Received: 26 July 1999 / Accepted: 22 November 1999 |
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