1887

Abstract

A modified gene from , encoding a variant of the green fluorescent protein (GFP), was subcloned into the mobilizable plasmid pMV158. was placed under the control of the inducible promoter of the gene , cloned in plasmid pLS70. The promoter is regulated by the product of the pneumococcal gene, which is inactivated by growing the cells in maltose-containing media. By homologous recombination, the construction was integrated into the host chromosome in a single copy. In both conditions (single and multiple copies), the pneumococcal cells were able to express GFP in an inducible or constitutive form, depending on whether the strain harboured a wild-type or a mutant gene. Quantification of the levels of GFP expressed by cultures supplemented with sucrose or maltose as carbon sources was feasible by fluorescence spectroscopy. Phase-contrast and fluorescence microscopy allowed pneumococcal cells expressing GFP in mixed cultures to be distinguished from those not carrying the gene.

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/content/journal/micro/10.1099/00221287-146-6-1267
2000-06-01
2020-09-25
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