Transformation of a type 4 encapsulated strain of Streptococcus pneumoniae

FEMS Microbiol Lett. 1999 Mar 15;172(2):131-5. doi: 10.1111/j.1574-6968.1999.tb13460.x.

Abstract

Streptococcus pneumoniae strain JNR.7/87 is a highly virulent, type 4 encapsulated Gram-positive bacterium whose transformability has not been tested previously, and whose genome is currently being sequenced. The strain was transformed at very low efficiency by addition of exogenous competence-stimulating peptide: However, the efficiency was too low and irreproducible to be useful in many genetic studies. Therefore, the effects on transformation efficiency of changing different components of competence-stimulating peptide-induced transformation have been examined. Screening of growth media was followed by optimization of pre-induction culture acidification, glycine concentration, and induction time. An optimized protocol was developed whereby S. pneumoniae strain JNR.7/87 was transformed reproducibly with a streptomycin resistance (SmR) marker at an efficiency of approximately 10(5) colony forming units per 10(8) cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / pharmacology
  • Culture Media / chemistry
  • DNA-Binding Proteins / pharmacology
  • Glycine
  • Humans
  • Streptococcus pneumoniae / genetics*
  • Streptococcus pneumoniae / growth & development
  • Time Factors
  • Transformation, Bacterial* / drug effects

Substances

  • Bacterial Proteins
  • ComA protein, Bacteria
  • Culture Media
  • DNA-Binding Proteins
  • Glycine