High-efficiency transformation and gene inactivation in Streptococcus suis type 2.
An efficient electrotransformation system for Streptococcus suis type 2 is described. It is demonstrated that vectors based on the broad-host-range plasmid pWVO1 replicate in S. suis type 2. Transformation efficiencies of about 10(7) transformants per micrograms of plasmid DNA could be obtained. Derivatives of plasmid pBR322 containing S. suis chromosomal DNA did not replicate but integrated into the chromosome. Southern hybridization analysis revealed that double as well as single cross-over integration events had occurred. Double cross-over events occurred at a frequency of about 15%. With these transformation and integration systems, recombinant DNA technology can now be applied to this important pathogenic species.
Smith
H E
HE
Department of Molecular Biology, Institute for Animal Science and Health, Lelystad, The Netherlands.
Wisselink
H J
HJ
Vecht
U
U
Gielkens
A L
AL
Smits
M A
MA
D015139:Blotting, Southern; D015153:Blotting, Western; D002876:Chromosomes, Bacterial; D003434:Crossing Over, Genetic; D004261:DNA Replication; D004269:DNA, Bacterial / Q000032:analysis / Q000096:biosynthesis; D004558:Electric Stimulation; D004797:Enzyme-Linked Immunosorbent Assay; D005798:Genes, Bacterial; D009154:Mutation; D010957:Plasmids; D015183:Restriction Mapping; D017011:Streptococcus suis / Q000235:genetics*; D014158:Transcription, Genetic; D014169:Transformation, Bacterial
D004269:DNA, Bacterial