A new vector-host system for construction of lacZ transcriptional fusions where only low-level gene expression is desirable.
We improved a multicopy vector, pRS415 [Simons et al., Gene 53 (1987) 85-96], for use in operon fusion constructions by introducing a new multiple cloning site (MCS) containing eight unique restriction sites upstream from the promoterless reporter gene lacZ. In order to reduce plasmid copy number, a new Escherichia coli strain SP2 (pcnB, delta lac, recA) was constructed. This strain permits analysis of fusions in cases where high gene dosage may be detrimental.
Podkovyrov
S M
SM
Department of Biochemistry and Anaerobic Microbiology, Virginia Polytechnic Institute and State University, Blacksburg 24061-0308, USA.
Larson
T J
TJ
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics*; D018628:Gene Dosage; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D007763:Lac Operon; D008969:Molecular Sequence Data; D009876:Operon / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000235:genetics
D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase