Improved vectors for nisin-controlled expression in gram-positive bacteria.
A set of shuttle vectors, able to replicate in Escherichia coli and in gram-positive bacteria, containing a nisin-inducible promoter (PnisA) and genes encoding NisR and NisK, the two-component signaling mechanism for activating transcription from PnisA in the presence of nisin, was constructed. To test these vectors, Enterococcus faecalis pCF10 plasmid genes prgX, prgY, and prgZ, which respectively encode cytosolic, integral membrane, and cell surface proteins, were cloned downstream of PnisA. Increased protein expression, in the presence of nisin, was demonstrated by Western blot analysis.
Bryan
E M
EM
Department of Microbiology, Institute for Advanced Studies in Biological Process Technology, Minneapolis, Minnesota 55455, USA.
Bae
T
T
Kleerebezem
M
M
Dunny
G M
GM
D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D017931:DNA Primers; D004261:DNA Replication; D013293:Enterococcus faecalis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005822:Genetic Vectors; D006094:Gram-Positive Bacteria / Q000187:drug effects / Q000235:genetics*; D008969:Molecular Sequence Data; D009561:Nisin / Q000378:metabolism* / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014157:Transcription Factors; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D017931:DNA Primers; D014157:Transcription Factors; C080935:nisR protein, Lactococcus lactis; D009561:Nisin