Construction of broad host range cloning vectors for gram-negative bacteria.

1 January 1985

journal article
Published by Oxford University Press (OUP) in Agricultural and Biological Chemistry

Vol. 49 (9) , 2719-2724
https://doi.org/10.1271/bbb1961.49.2719

Abstract

A new cloning vector, pMFY31, has been constructed based on the high-copy-number, broad-host-range plasrnid RSF1010. The plasmid has a size of 13.2kb and carries the Ap^r, Cm^r, and Tc^r genes. It contains unique PstI, EcoRI, HindIII, BamHI, and SalI sites, all of which are located within the antibiotic resistance genes, therefore all sites are applicable to insertional inactivation. We also constructed pMFY40, a 11.6 kb derivative of pMFY31, by the elimination of the Cm^r gene. Plasmid pMFY31 has been efficiently introduced into a Pseudomonas putida strain not only by plasmid-DNA transformation but also by conjugal co-transfer with the helper plasmid, and was maintained stably in the strain.

Keywords

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