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Potential for gene transfer between manipulated bacteria and the resident microflora of the human gut. - FS0207

1.To express lux and gfp marker genes in at least one strain of Lactococcus lactis and Enterococcus faecalis. 2.To introduce lux and/or gfp marker genes into the chromosome of L. lactis. 3.To establish fermenter systems modelling the human hind gut. 4.To establish and validate procedures for detecting lux and gfp gene sequences in DNA extracted from mixtures of human gut microorganisms and added, marked strains, by PCR amplification. 5.To develop fluorimetric procedures for detecting and enumerating gfp- marked lactic acid bacteria added to mixed human gut microorganisms. 6.To follow the fate of gfp-marked lactic acid bacteria and gfp and lux gene sequences introduced into fermenters that simulate conditions in the human gut; to screen for transfer to endogenous microflora. 7.To examine the stability of radiolabelled chromosomal DNA from Lactococcus lactis to gut conditions. 8.To study the influence of gut factors upon transfer of plasmids and chromosomal elements between Lactococcus strains and endogenous lactic acid bacteria from the gut. 9.To investigate acquisition of genetic elements by introduced lactic acid bacteria from endogenous microbes.
Time-Scale and Cost
From: 1995

To: 1998

Cost: £235,067
Contractor / Funded Organisations
Rowett Research Institute