Part:BBa_K4347006

Sac7e

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].

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.

DNA binding protien "Sac7e" modelled in Pymol.

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
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References

1. https://www.nature.com/articles/srep37274

2. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC373405/