New gentamicin-resistance and lacZ promoter-probe cassettes suitable for insertion mutagenesis and generation of transcriptional fusions.
A set of antibiotic-resistance and promoter-probe cassettes suitable for insertion mutagenesis and generation of transcriptional fusions was constructed. The cassettes contain the aacC1 gene of transposon Tn1696 conferring resistance to gentamicin in a large variety of Gram- and Gram+ bacteria. In addition to the antibiotic-resistance gene, a promoterless Escherichia coli lacZ gene was included in the cassettes, allowing the determination of the transcriptional activity at the insertion site. The cassettes can be excised from a plasmid mediating ampicillin resistance by many commonly used restriction enzymes. The new constructs have been successfully used for mutagenesis and studies of gene transcription in Rhizobium meliloti.
Becker
A
A
Lehrstuhl für Genetik, Fakultät für Biologie, Universität Bielefeld, Germany.
Schmidt
M
M
Jäger
W
W
Pühler
A
A
D000123:Acetyltransferases / Q000235:genetics; D001483:Base Sequence; D004352:Drug Resistance, Microbial / Q000235:genetics; D005819:Genetic Markers; D005839:Gentamicins / Q000494:pharmacology; D007763:Lac Operon; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000379:methods*; D012313:RNA / Q000096:biosynthesis*; D012641:Selection, Genetic; D014158:Transcription, Genetic
D005819:Genetic Markers; D005839:Gentamicins; C039919:RNA, recombinant; D012313:RNA; D000123:Acetyltransferases; C020564:gentamicin 3-acetyltransferase