Part:BBa_K4347011

Bst fusion with Sac7e and point mutations for enhanced thermal stability codon optimized for E.coli

This fusion protien was designed by linking the N-terminus of a modified Bst polymerase with thermostable DNA binding protien Sac7e using a flexible (GGGGS)₄ linker to increase polymerase thermostability and processivity in LAMP reaction.

Usage and Biology

Usage of bst and Sac7e in biology. Can summarize what was written for "bst with point mutations" and "sac7e" pages. This bst version DOES include the point mutations

A more thermally stable and processive polymerase

This final iteration of the new polymerase is an improvement of our previous part; BBa_K4347010, as it is a combination of our more thermally stable polymerase (BBa_K4347007) fused with DNA binding protien Sac7e (BBa_K4347006).

Small summary on the new polymerase, combining parts https://parts.igem.org/Part:BBa_K4347006 and https://parts.igem.org/Part:BBa_K4347007#

Thermal stability of Bst fusion protien computed from YASARA.

Lab results and pictures

Assembly Compatibility:

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

  Illegal BamHI site found at 5
  Illegal XhoI site found at 209
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1015
  
• 1000
  COMPATIBLE WITH RFC[1000]