Difference between revisions of "Part:BBa K4347006"
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===Usage and Biology=== | ===Usage and Biology=== | ||
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| + | [[File:BBa K4347006 sac7e.PNG|300px|left|thumb|DNA binding protien "Sac7e" modelled in Pymol.]] | ||
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[[Part:BBa_K4347006#References|<sup>[1]</sup>]]. These protiens fold as an SH3-like domain capped by a C-terminal α-helix[[Part:BBa_K4347006#References|<sup>[2]</sup>]] and they sharply kink the double DNA helix upon binding into the minor groove[[Part:BBa_K4347006#References|<sup>[3]</sup>]]. | 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[[Part:BBa_K4347006#References|<sup>[1]</sup>]]. These protiens fold as an SH3-like domain capped by a C-terminal α-helix[[Part:BBa_K4347006#References|<sup>[2]</sup>]] and they sharply kink the double DNA helix upon binding into the minor groove[[Part:BBa_K4347006#References|<sup>[3]</sup>]]. | ||
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Compared to other similar proteins, Sac7e showed the highest affinity for dsDNA (KD = 11 μM), with binding sites ~ 6-8 bases per protein. | Compared to other similar proteins, Sac7e showed the highest affinity for dsDNA (KD = 11 μM), with binding sites ~ 6-8 bases per protein. | ||
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Revision as of 18:25, 18 July 2022
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]. These protiens fold as an SH3-like domain capped by a C-terminal α-helix[2] and they sharply kink the double DNA helix upon binding into the minor groove[3].
Previous work shows that fusing Sso7d (Sac7e homologue) to Taq polymerase (Bst polymerase homologue) improves processivity and improves PCR amplification[4]. We hope to use the more thermostable homologues to create a new fusion protien that will improve processivity in LAMP amplification.
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
1. https://www.nature.com/articles/srep37274
2. https://pubmed.ncbi.nlm.nih.gov/7577913/