pHG165: A pBR322 copy number derivative of pUC8 for cloning and expression |
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| Authors: | Gordon S. A. B. Stewart Sharon Lubinsky-Mink Clive G. Jackson Aliza Cassel Jonathan Kuhn |
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| Affiliation: | 1. Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER), Universidad de Sevilla-CSIC-Universidad Pablo de Olavide, Sevilla, Spain;2. Departamento de Genética, Universidad de Sevilla, Sevilla, Spain;3. Institute of Molecular Biology (IMB), Mainz, Germany |
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| Abstract: | During the construction of the Messing pUC plasmid series, the rop(rom) gene of pBR322 which mediates the activity of RNAI was deleted. This has resulted in an elevated copy number for the pUC plasmids which makes the expression of beta-galactosidase activity constitutive in a host containing the Iqtss lac repressor. We describe the construction of a new series of vectors which retain the pUC multiple cloning site (MCS) but in which copy number control has been recovered. In addition, the lac alpha/lac promoter expression region has been inserted into a HpaI cassette. This facilitates the movement of recombinant DNA clones within the MCS. It also increases the complementation activity of the lac alpha peptide by an order of magnitude, allowing selection of recombinants by their Lac- phenotype on MacConkey agar. |
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