Genetic analysis of the gentamicin resistance region of pPH1JI and incorporation into a wide host range cloning vehicle.
A region of the IncP plasmid pPH1JI encoding resistance to gentamicin, spectinomycin, and streptomycin was characterized by subcloning, deletion, and insertion mutagenesis. Approximate locations of these resistance determinants were established. A 1.6-kb HindIII-SphI segment of this region expresses gentamicin resistance (Gmr) in Escherichia coli when inserted into various plasmid vectors; this DNA segment encodes a polypeptide of 17.5 kDa. Incorporation of this fragment into an IncP cloning vehicle produced a Gmr wide host range vector, pRAR209, which confers levels of Gmr comparable to those expressed by pPH1JI in E. coli, Agrobacterium tumefaciens, and Rhizobium meliloti.
Rubin
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BioTechnica International, Inc., Cambridge, Massachusetts 02140.
D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D005839:Gentamicins / Q000494:pharmacology*; D010957:Plasmids; D011815:R Factors
D005839:Gentamicins; D004262:DNA Restriction Enzymes