Transformation of Helicobacter Pylori
Categorized: Bacterial Genetics
O’Toole et al, 1994, Mol. Microbiol. 14: 691-703.
- Plate-grown H. pylori are resuspended in BHI to an OD600 of 0.6.
- 1 µg of DNA is added and incubated for 3 h at 37ºC (in a CO2 incubator).
- The cells are then spun down and resuspended in 100ul of BHI.
- The entire 100 µl is spread onto a chocolate blood agar (CBA) plate (no antibiotic) and incubated for 24 h.
- After 24 h the cells are scraped or washed from the plate and replated on CBA with antibiotic. Single colonies are restreaked for further analysis.
- PCR can then be used to determine if the gene replacements have occurred as predicted and to confirm that no other rearrangements have occurred.