The use of bacterial luciferase genes as reporter genes in Lactococcus: regulation of the Lactococcus lactis subsp. lactis lactose genes.
Lactose metabolism is an important industrial trait in dairy lactococci. In Lactococcus lactis, lactose is taken up via the phosphoenolpyruvate-dependent phosphotransferase system (PEP-PTS) and is subsequently metabolized via the glycolytic and tagatose 6-phosphate pathways. Genes for the lactose-specific PEP-PTS proteins, phospho-beta-galactosidase and tagatose 6-phosphate pathway enzymes are encoded by a single 8 kb operon, lacABCDFEGX, and there is a divergently transcribed lacR repressor gene. Transcriptional fusions of both the lac operon promoter and the lacR promoter to the luxAB genes of Vibrio fischeri were used to investigate the regulation of expression of both promoters. In vivo bioluminescence assays demonstrated that lacR negatively regulates the lac operon and also autoregulates itself. Induction of transcription occurred for both promoters during growth on lactose: sevenfold for lacR and fivefold for the lac operon. The lacR promoter was demonstrated to be a particularly strong promoter, being approximately four times more efficient than the lac operon promoter. Both promoters provide good potential for the inducible expression of foreign proteins in Lactococcus.
AFRC Institute of Food Research, Colney, Norwich, UK.
T J
TJ
Eaton
C A
CA
Shearman
M J
MJ
Gasson
D001483:Base Sequence; D004790:Enzyme Induction; D015964:Gene Expression Regulation, Bacterial; D007763:Lac Operon / Q000235:genetics*; D013294:Lactococcus lactis / Q000235:genetics*; D007785:Lactose / Q000378:metabolism; D008156:Luciferases / Q000235:genetics; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics*; D014158:Transcription, Genetic; D014733:Vibrio / Q000201:enzymology / Q000235:genetics
D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D008156:Luciferases; D007785:Lactose