Part:BBa_K4347006
Sac7e
Contents
Sac7e is a DNA binding protien from the “7 kDa DNA-binding” family which is composed of chromatin proteins from the Sulfolobales archaeal order. Sac7e was isolated from thermoacidophilic archaeon Sulfolobus acidocaldarius and is structurally homologous to DNA binding protien Sso7d isolated from thermophilic archaeon Sulfolobus solfataricus [1].
Structural homologue Sso7d from 2021 Fudan team: https://parts.igem.org/Part:BBa_K3790002
Usage and Biology
DBA binding protiens from the 7 kDa DNA-binding family are highly thermostable and resistant to extreame pH conditions. These protiens have high affinity for double stranded DNA, thus their biological role is to bind genomic DNA in order to prevent its melting at the high growth temperatures of thermophilic archaea[1].
Previous work shows that fusing Sso7d (Sac7e homologue) to Taq polymerase (Bst polymerase homologue) improves processivity and improves PCR amplification[2]. We hope to use more thermally stable homologues to create a new fusion protien that will improve processivity in LAMP amplification.
These proteins are hyper-thermostable and resistant to pH extremes. Sac7e is thermally stable to 85.5°C.
Compared to other similar proteins, Sac7e showed the highest affinity for dsDNA (KD = 11 μM), with binding sites ~ 6-8 bases per protein.
- 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]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
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