Construction of broad-host-range vectors for general cloning and promoter selection in Pseudomonas and Escherichia coli |
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Authors: | M A Farinha A M Kropinski |
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Institution: | Department of Microbiology and Immunology, Queen's University, Kingston, Ont., Canada. |
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Abstract: | We have constructed two promoter-selection vectors based upon the broad-host-range plasmid pRO1614. pQF40 (6 kb) contains a promoterless tetA gene downstream from a large multiple cloning site while pQF26 (5.4 kb) possesses a promoterless cat cartridge. The latter vector displayed a copy number of 13 in Pseudomonas aeruginosa and 39 in Escherichia coli. When promoter sequences derived from the Pseudomonas phage phi PLS27 were cloned into pQF26, high levels of chloramphenicol-acetyltransferase were detected in P. aeruginosa. In E. coli the activity was approximately one-third that in P. aeruginosa when corrections were made for the plasmid copy number. |
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