Difference between revisions of "Part:BBa K4347007"
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===Usage and Biology===
 
===Usage and Biology===
  
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Bst polymerase is a family I DNA polymerase derived from the thermophilic bacterium <em> Geobacillus stearothermophilus.</em> Bst polymerase Large Fragment is notable for its strong strand displacement activity and thermal stability [[Part:BBa_K4347007#References|<sup>[1]</sup>]]. Bst also contains a 5' to 3' DNA polymerase activity but lacks 3' to 5' exonuclease activity [[Part:BBa_K4347007#References|<sup>[2]</sup>]]. These unique features of Bst make it
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Bst polymerase is a family I DNA polymerase derived from the thermophilic bacterium <em> Geobacillus stearothermophilus.</em> Bst polymerase Large Fragment is notable for its strong strand displacement activity and thermal stability [[Part:BBa_K4347007#References|<sup>[1]</sup>]]. Bst also contains a 5' to 3' DNA polymerase activity but lacks 3' to 5' exonuclease activity [[Part:BBa_K4347007#References|<sup>[2]</sup>]]. These unique features allow Bst polymerase to facilitate isothermal amplification techniques such as LAMP and rt-LAMP.
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===Enhanced Thermal stability===
 
===Enhanced Thermal stability===

Revision as of 15:35, 11 July 2022

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 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.



Enhanced Thermal stability

Talk about the point mutations. Pictures of lab experiments


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