In vivo formation of gene fusions in Pseudomonas putida and construction of versatile broad-host-range vectors for direct subcloning of Mu d1 and Mu d2 fusions.
The Mu d1 and Mu d2 prophages were integrated into the conjugative broad-host-range plasmid R751. The two plasmids were then transferred into Pseudomonas putida, and derivatives carrying intact Mu prophages were recovered. After induction of Mu at 42 degrees C, both operon and gene fusions were observed on 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal) plates. Broad-host-range vectors were constructed which allow direct cloning of both operon or gene fusions and their analysis in Escherichia coli and P. putida. By using one of these vectors, two operon fusions were isolated from the P. putida chromosome and comparatively analyzed in E. coli and P. putida.
Simon
V
V
Lehrstuhl für Genetik, Universität Bayreuth, Federal Republic of Germany.
Schumann
W
W
D001435:Bacteriophages / Q000235:genetics*; D003001:Cloning, Molecular; D003227:Conjugation, Genetic; D004262:DNA Restriction Enzymes; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D015245:Deoxyribonuclease BamHI; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005822:Genetic Vectors; D009693:Nucleic Acid Hybridization; D009876:Operon; D010957:Plasmids; D014176:Protein Biosynthesis; D011549:Pseudomonas / Q000235:genetics*; D014158:Transcription, Genetic
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI