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Volume 146, Issue 6

Quantitative detection of cells harbouring single or multiple copies of the gene encoding the green fluorescent protein Free

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.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): Ap, ampicillin , chromosomal integration , Cm, chloramphenicol , GFP, green fluorescent protein , green fluorescent protein , MalR repressor , maltose regulon , pneumococci , Tc, tetracycline and TIR, translation initiation region
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2000-06-01
2024-04-23
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Quantitative detection of Streptococcus pneumoniae cells harbouring single or multiple copies of the gene encoding the green fluorescent protein
Microbiology 146, 1267 (2000); https://doi.org/10.1099/00221287-146-6-1267
/content/journal/micro/10.1099/00221287-146-6-1267
/content/journal/micro/10.1099/00221287-146-6-1267
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