Cloning of the lux genes into Lactobacillus casei and Streptococcus lactis: phosphate-dependent light production

Ahmad, Khalid A.; Stewart, Gordon S. A. B.

doi:10.1042/bst0161068

Cited by 15 publications

(7 citation statements)

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“…As it is not possible to clone proU promoter-up mutations on multicopy plasmids, the low copy number plasmid pHSG575 was used (Takeshita et al, 1987). The lux reporter genes from pSB71 (Ahmad and Stewart, 1988) were cloned into pHSG575 as a 2 kb EcoRI-BamHI fragment. The wild-type, proU1718 and proU1719 promoters, extending from base pairs -217 to +100, were cloned upstream of the lux gene as 317 bp EcoRI fragments, amplified by the PCR from chromosomal DNA of strains CH1512, CH1973 and CH 1974, respectively.…”

Section: Sequences Of the Mutant Promotersmentioning

confidence: 99%

“…The low copy number plasmid pHSG575 (Takeshita et al, 1987) was digested with EcoRI and BamHI and the vector fragment purified from an agarose gel. Plasmid pSB71 (Ahmad and Stewart, 1988) was also digested with EcoRI and BamHI, the 2 kb fragment containing the lux reporter genes from Vibrio fischeri isolated and cloned into pHSG575.…”

Section: Plasmid Construction and Site-directed Mutagenesismentioning

confidence: 99%

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DNA twist, flexibility and transcription of the osmoregulated proU promoter of Salmonella typhimurium.

et al. 1995

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Transcription from many bacterial promoters is sensitive to the level of DNA supercoiling. We have investigated the mechanism by which environmentally induced changes in DNA supercoiling might regulate transcription. For the proU promoter of Salmonella typhimurium, osmotically induced changes in DNA topology appear to play a primary regulatory role. Changes in DNA supercoiling (linking number; delta Lk) are partitioned into changes in the winding of the strands of the double helix about themselves (twist; delta Tw) and/or elastic deformations or flexibility of the DNA helix (writhe; delta Wr). Mutations of the proU promoter were isolated in vivo, or generated in vitro, which altered the spacing between the −10 and −35 motifs. Studies on these mutant promoters, both in vivo and in vitro, exclude models in which changes in DNA twist play a regulatory role. Instead, our data suggest that increased DNA flexibility, reflecting the osmotically induced increase in negative supercoiling of DNA, is required for promoter activation.

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Section: Sequences Of the Mutant Promotersmentioning

confidence: 99%

Section: Plasmid Construction and Site-directed Mutagenesismentioning

confidence: 99%

DNA twist, flexibility and transcription of the osmoregulated proU promoter of Salmonella typhimurium.

et al. 1995

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“…A promoterless copy of the Vibrio$scheri luxAB genes was then introduced into this backbone. The plasmid pSB 185 (Ahmad & Stewart, 1988) contains a promoterless copy of the luxAB genes but retains the native ShineDalgarno sequence (Fig. 1).…”

Section: Construction Of a Broad Host Range Promoter Probe Plasmidmentioning

confidence: 99%

“…Additionally, PrfA is known to co-ordinately regulate the expression of other genes in the virulence chromosomal region (Vazquez-Boland et al, 1992;Chakraborty et al, 1992), the products of which include metalloprotease 'wpl), lecithinase (plcB) and an actin nucleation factor BctA). Gasson & Anderson (1985); 4, Ahmad & Stewart (1988).…”

Section: Introductionmentioning

confidence: 99%

The use of bacterial luciferase for monitoring the environmental regulation of expression of genes encoding virulence factors in Listeria monocytogenes

Park

Stewart

Kroll

1992

Journal of General Microbiology

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A promoter probe vector, which utilized the luxAB genes from Vibriofischeri as reporters of gene expression, was constructed for use in Listeriu monocytogenes. Using this system gene expression can be monitored nondestructively and in real-time, simply by measuring cellular bioluminescence. Derivatives of the promoter probe were constructed that contained the cloned promoters from the hlyA and plcA genes of L. monocytogenes. The activity of these promoters was dependent on the transcriptional activator PrfA. Accordingly, in a strain containing an intact copy of the prfA gene, expression from both the hlyA and plcA promoters was 25-45-fold higher than inprfA mutants. Heat shock was identified as an environmental signal which induced expression of hlyA and plcA. Conversely, oxidative stress had no effect upon the expression of the virulence factors. In addition, the composition of the growth media was found to have a dramatic effect upon the expression of @ A and plcA, suggesting the presence of an unidentified signal which may regulate induction of expression of virulence genes in L. monocytogenes.

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“…lux Expression elements functional in lactic acid bacteria have been con w structed to provide novel reagents for the detection of antimicrobial activity in milk (Ahmad and Stewart, 1988). Inhibition of starter culture bacteria may be caused by antibiotics, detergents~disinfectants or bacteriophage, and although, in many cases, the precise identification of the inhibitory substance is not required, there is a need for a test which reports their presence more rapidly than the current and extensively used disc and dye reduction assays (Harrigan and McCance, 1976).…”

Section: Antibiotic Detectionmentioning

confidence: 99%

Biotechnology-Based Methods for the Detection, Enumeration and Epidemiology of Food Poisoning and Spoilage Organisms

Dodd

Stewart

Waites

1990

Biotechnology and Genetic Engineering Reviews

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Cloning of the lux genes into Lactobacillus casei and Streptococcus lactis: phosphate-dependent light production

Cited by 15 publications

References 1 publication

DNA twist, flexibility and transcription of the osmoregulated proU promoter of Salmonella typhimurium.

DNA twist, flexibility and transcription of the osmoregulated proU promoter of Salmonella typhimurium.

The use of bacterial luciferase for monitoring the environmental regulation of expression of genes encoding virulence factors in Listeria monocytogenes

Biotechnology-Based Methods for the Detection, Enumeration and Epidemiology of Food Poisoning and Spoilage Organisms

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