A simple and efficient system for the construction of phoA gene fusions in gram-negative bacteria.
We have developed a two-plasmid system for generating gene fusions between phoA and cloned genes encoding envelope proteins. The vector plasmid carries a temperature-sensitive replication system and can be rescued at high temperature by insertion of an IS1-based transposon carrying the ori region of pBR322 and a phoA gene lacking transcription and translation initiation signals. The vector plasmid also carries the transfer origin of the conjugative plasmid, F, permitting transfer into a suitable recipient strain. We have used this system in the analysis of the bla gene cloned from pBR322.
Duchêne
A M
AM
Molecular Genetics and Microbiology Unit (CNRS), Toulouse, France.
Patte
J
J
Gutierrez
C
C
Chandler
M
M
D000469:Alkaline Phosphatase / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001483:Base Sequence; D004261:DNA Replication / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D013696:Temperature; D001618:beta-Lactamases / Q000235:genetics*
D001425:Bacterial Outer Membrane Proteins; D004251:DNA Transposable Elements; D011993:Recombinant Fusion Proteins; D000469:Alkaline Phosphatase; D001618:beta-Lactamases