pUdOs: concise plasmids for bacterial and mammalian cells

Ourida Manigat, F.; Connell, L. B.; Stewart, B. N.; LePabic, A.-R.; Tessier, C. J. G.; Emlaw, J. R.; Calvert, N. D.; Rossl, A.; Shuhendler, A. J.; daCosta, C. J. B.; Campbell-Valois, F.-X.

doi:10.1101/2023.07.05.547852

bioRxiv · 10.1101/2023.07.05.547852

pUdOs: concise plasmids for bacterial and mammalian cells

Ourida Manigat, F.·Connell, L. B.·Stewart, B. N.·LePabic, A.-R.·Tessier, C. J. G.·Emlaw, J. R.·Calvert, N. D.·Rossl, A.·Shuhendler, A. J.·daCosta, C. J. B.·Campbell-Valois, F.-X.

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Abstract

The pUdOs are 28 plasmids of small size combining four different origins of replication and seven selection markers, which together afford flexible use in Escherichia coli and several related gram- negative bacteria. The promoterless multicloning site is insulated from upstream spurious promoters by strong transcription terminators, and contains type IIP or IIS restriction sites for conventional or Golden-gate cloning. pUdOs can be converted into efficient expression vectors through the insertion of a promoter at the users discretion. For example, we demonstrate the utility of pUdOs as the backbone for an improved version of a Type III Secretion System reporter in Shigella. In addition, we derive a series of pUdO-based mammalian expression vectors affording distinct levels of expression and transfection efficiencies comparable to commonly used mammalian expression plasmids. Thus, pUdOs could advantageously replace traditional plasmids in a wide variety of cell types and applications.

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Ourida Manigat, F., Connell, L. B., Stewart, B. N., LePabic, A.-R., Tessier, C. J. G., Emlaw, J. R., Calvert, N. D., Rossl, A., Shuhendler, A. J., daCosta, C. J. B., Campbell-Valois, F.-X.. 2023-07-05. pUdOs: concise plasmids for bacterial and mammalian cells. https://doi.org/10.1101/2023.07.05.547852

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