Table 1.
Methods of competency and transformation for selected thermophilic prokaryotes of commercial relevance.
| Organism | Genome sequence | Competency method | Transformation method | Maximum reported efficiency | Reference |
|---|---|---|---|---|---|
| Thermoanaerobacterium saccharolyticum JW/SL‐YS485 | Department of Energy (DOE) Joint Genome Institute | Autoplast generation with cell wall‐destabilizing agent (niacin), subsequent sucrose‐containing buffer washes | Electroporation (12.5 kV cm−1, 400 ohm, 25 µF) | Approximately 103–4 | Tyurin et al. (2006), Shaw et al. (2008) |
| Thermoanaerobacter ethanolicus JW200 | DOE Joint Genome Institute | Glycine‐ and sucrose‐induced protoplast formation, subsequent glycerol‐containing buffer washes | Electroporation (13 kV cm−1, 400 ohm, 25 µF) | Approximately 101 | Peng et al. (2006) |
| Thermoanaerobacter mathranii BG1 | Not recorded | Cellobiose wash buffer. Isoniacin addition | Custom built electroporator with custom cuvettes (2mm gap). 10 ms square wave pulse. 25 kV cm−1 | 102−5 | Yao and Mikkelsen (2010) |
| Geobacillus thermoglucosidasius NCIMB 11955 | Not recorded | High osmolarity washing buffer (sorbitol and mannitol) | Electroporation (25 kV cm−1, 600 ohm, 10 µF) | 104 | Taylor et al. (2008), Cripps et al. (2009) |
| Geobacillus thermoglucosidasius DL33 | Not recorded | High osmolarity washing buffer (sorbitol and mannitol) | Electroporation (25 kV cm−1, 600 ohm, 10 µF) | 103−4 | Taylor et al. (2008), Cripps et al. (2009) |
| Clostridium thermocellum | DOE Joint Genome Institute | Isoniacin addition | Custom built electroporator with custom cuvettes (2mm gap). 10 ms square wave pulse. 25 kV cm−1 | 103 | Tyurin et al. (2004) |
| Rhodothermus marinus (trpB‐) | DOE Joint Genome Institute | Glycerol treatment | Electroporation (22.5 kV cm−1, 200 ohm, 25 µF) | 106 | Bjornsdottir et al (2005; 2006; 2007) |
| Thermosynechococcus elongatus BP‐1 | Kasuza DNA Research Institute | 2mM N‐tris(hydroxymethyl)methyl‐2‐aminoethanesulfonic acid (TES) treatment | Electroporation (10 kV cm−1, 200 ohm, 25 µF) and natural competency | 109 | Iwai et al. (2004) |
| Thermotoga maritima, T. neapolitana | J. Craig Venter Institute | Lysozyme treatment and DNA encapsulation with liposome reagent | Spheroplast creation and cationic liposome integration | 102 | Noll and Vargas (1997), Yu et al. (2001), Conners et al. (2006) |
| Thermus thermophilus HB27 | Goettingen Genomics Laboratory | Glycerol treatment | Natural competency and electroporation (25 kV cm−1) | 109 | Friedrich et al. (2001) |
| Pyrococcus furiosus DSM 3638 | Universities of Utah and Maryland | CaCl2 treatment | Heat shock at 80°C | 102 | Waege et al. (2010) |
| Thermococcus kodakaraensis KOD1 | Kyoto University | CaCl2 treatment | Heat shock at 85°C | 102 | Fukui et al. (2005), Sato et al. (2005) |
| Sulfolobus islandicus, S. solfataricus | University of Copenhagen, DOE Joint Genome Institute | Sucrose treatment in growth medium and wash buffers | Electroporation (7.5 kV cm−1, 400 ohm, 25 µF) | 103 | She et al. (2001; 2009), Albers and Driessen (2008) |