Difference between revisions of "Part:BBa K4347011"
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===A more thermally stable and processive polymerase=== | ===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). | + | 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). The modified Bst polymerase contains three point mutations in the polymerase thumb domain: K549W, K582L and Q584L, which have been proven to improve thermal stability |
Small summary on the new polymerase, combining parts https://parts.igem.org/Part:BBa_K4347006 and https://parts.igem.org/Part:BBa_K4347007# | Small summary on the new polymerase, combining parts https://parts.igem.org/Part:BBa_K4347006 and https://parts.igem.org/Part:BBa_K4347007# | ||
Revision as of 17:07, 20 July 2022
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). The modified Bst polymerase contains three point mutations in the polymerase thumb domain: K549W, K582L and Q584L, which have been proven to improve thermal stability
Small summary on the new polymerase, combining parts https://parts.igem.org/Part:BBa_K4347006 and https://parts.igem.org/Part:BBa_K4347007#
Lab results and pictures
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 5
Illegal XhoI site found at 209 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1015
- 1000COMPATIBLE WITH RFC[1000]