New gentamicin-resistance and lacZ promoter-probe cassettes suitable for insertion mutagenesis and generation of transcriptional fusions
1995; Elsevier BV; Volume: 162; Issue: 1 Linguagem: Inglês
10.1016/0378-1119(95)00313-u
ISSN1879-0038
AutoresAnke Becker, Michael Schmidt, Wolfgang Jäger, Alfred Pühler,
Tópico(s)Bacterial Genetics and Biotechnology
ResumoA set of antibiotic-resistance and promoter-probe cassettes suitable for insertion mutagenesis and generation of transcriptional fusions was constructed. The cassettes contain the aacCl 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.
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