Part:BBa_K4347007

Bst with point mutations for enhanced thermal stability codon optimized for E.coli

This part is an improvement from Bst Polymerase 1 (large fragment) for E.coli from the 2021 iGEM Fudan team: https://parts.igem.org/Part:BBa_K3790000.

Usage and Biology

Bst polymerase Large Fragment is a family I DNA polymerase derived from the thermophilic bacterium Geobacillus stearothermophilus. Bst polymerase Large Fragment is notable for its strong strand displacement activity and thermal stability ^[1]. Bst also contains a 5' to 3' DNA polymerase activity but lacks 3' to 5' exonuclease activity ^[2]. These unique features allow Bst polymerase to facilitate isothermal amplification techniques such as LAMP and rt-LAMP.

Bst polymerase derived from thermophilic bacterium Geobacillus stearothermophilus used in LAMP modelled on PyMol.

Enhanced Thermal stability

The origional codon optimized Bst polymerase (Part BBa_K3790000) was improved upon to further enhance the polymerases thermal stability such that it could function at a higher temperature.

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 766
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

1. https://www.tandfonline.com/doi/full/10.1080/10826068.2022.2095573?src=

2. https://pubmed.ncbi.nlm.nih.gov/8740835/ Aliotta JM, Pelletier JJ, Ware JL, Moran LS, Benner JS, Kong H. Thermostable Bst DNA polymerase I lacks a 3'-->5' proofreading exonuclease activity. Genet Anal. 1996 Mar;12(5-6):185-95. PMID: 8740835