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TmaSSB Protein

 TmaSSB Protein
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Extreme Thermostable Single-Stranded DNA Binding Protein isolated from the hyperthermophilic bacteria Thermotoga neapolitana (TneSSB). Due to the extreme thermostability, TneSSB can be used in molecular biology applications that require extremely high temperature conditions, such as nucleic acid amplification and sequencing.
 
Thermostability
TneSSB is the most thermostable SSB protein identified to date, with a melting temperature (Tm) of 112.5°C  (Tm of TaqSSB is 86.8°C). The half-live of the ssDNA-binding activity at 100°C is 12 h.
 Extreme Thermostable Single-Stranded DNA Binding Protein isolated from the hyperthermophilic bacteria Thermotoga maritima (TmaSSB). Due to the extreme thermostability, TmaSSB can be used in molecular biology applications that require extremely high temperature conditions, such as nucleic acid amplification and sequencing.
 
Thermostability
TmaSSB is an extreme thermostable SSB protein, with a melting temperature (Tm) of 109.3°C (Tm of TaqSSB is 86.8°C). The half-live of the ssDNA-binding activity at 100°C is 10 h.
 
Applications of thermostable SSB proteins:
- Prevents PCR inhibition (2)
- Increases amplification efficiency (3,4,5,6,9)
- Increases selectivity and specificity of multiplex PCR (7)
- Protects single stranded DNA from degradation (8)
- Reduces secondary structure formation, which inhibits PCR
- Enhances amplification of, so called, difficult templates (e.g. rich in GC)
- Stimulates fidelity and processivity of Taq polymerase (9)
- Reacts with RNA allowing increase of a synthesized cDNA size (9)
- Stabilises ssDNA in the site-specific mutagenesis
- Facilitate in obtaining complete digestion by restriction endonucleases
 
Product Source
E. coli strain that carries the cloned ssb gene from Thermotoga maritima.
 
Storage buffer
20 mM Tris-HCl (pH 8.0), 300 mM NaCl, 1 mM EDTA and 50% (v/v) glycerol
 
Size
250 µg
 
Concentration
1 mg/ml
 
Storage conditions
store at-20°C.
 
Quality control:
Lack of endonuclease and exonuclease activity. Electrophoretic purity over 95% (SDS-PAGE).
 
Note
Add 0.2-0.5 µg (0.4-1 µl) of TmaSSB per 50 µl reaction.
 
References
1. Greipel J., Urbanke C., and Maass G., Modulation of the affinity of the single-stranded DNA-binding protein of Escherichia coli (E. coli SSB) to poly(dT) by site-directed mutagenesis.  in: Saenger, W., Heinemann, U. (Eds.) pp. 61-86 (1989)
2. Kreader C., Relief of amplification inhibition in PCR with bovine serum albumin or T4 gene 32 protein. Applied Environ. Micro. 62, 1102-1106 (1996)
3. Dąbrowski S., Olszewski M., Piątek R. and Kur J., Novel thermostable ssDNA-binding proteins from Thermus thermophilus and T. aquaticus-expression and purification.  Protein Expr. Purif. 26, 131-138 (2002)
4. Dąbrowski S. and Kur J., Cloning, overexpression, and purification of the recombinant His-tagged SSB protein of Escherichia coli and use in polymerase chain reaction amplification.  Protein Expr. Purif. 16, 96-102 (1999)
5. Rapley R., Enhancing PCR amplification and sequencing using DNA-binding proteins. Mol. Biotech. 2, 295-298 (1994)
6. Schwarz K., Hansen-Hagge T. and Bartram C., Improved yields of long PCR products using gene 32 protein. Nucleic Acids Res. 18, 1079 (1990)
7. Barski P., Piechowicz L., Galinski J. and Kur J., Rapid assay for detection of methicillin-resistant Staphylococcus aureus using multiplex PCR. Mol. Cell Probes 10, 471-475 (1996)
8. Fradkin G, Aizenberg O, Torosian M, Shishkova O, Enzymatic mechanisms of degradation of DNA replication forks in vitro. Mol Biol. 18, 1518-23 (1984)
9. Perales C., Cava F., Meijer W., Berenguer J., Enhancement of DNA, cDNA synthesis and fidelity at high temperatures by a dimeric single-stranded DNA-binding protein.  Nucleic Acids Res. 31, 6473-6480 (2003)
 
 
We own a substantial collection of thermostable SSB proteins:
- TaqSSB (from Thermus aquaticus)
- TthSSB (from Thermus thermophilus)
- DraSSB (from Deinocossus radiopugnans)
- DgeSSB (from Deinocossus geothermalis)
- DmuSSB (from Deinocossus murrai)
- TneSSB (from Thermotoga neapolitana)
- TmaSSB (from Thermotoga maritima)
- TteSSB (from Thermoanaerobacter tengcongensis)
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