Part:BBa_K4439010

GST-Tag

Abstract

There are multiple protein affinity tags that are being used for protein purification and the GST-tag is one of them. GST is short for Glutathione-S-transferase and refers to a whole protein. The GST enzyme tag has been in use for protein purification since the late 1980s (Smith & Johnson 1988) and has been established as a reliable way for protein purification assays that require the highest level of purity (Harper & Speicher 2010).

The GST-tag size is 26 kDa and therefore really large compared to other protein affinity tags. In theory, it can be fused to either the C- or N-terminus of a protein (Terpe 2003), but we highly recommend adding it to the N-terminus. So far it has been successfully used to purify proteins from bacterial cells (Smith and Johnson 1988), yeast (Lu et al. 1997), and mammalian cells (Rudert et al. 1996).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 82
  

References

• Smith, D. B. & Johnson, K. S. (1988, juillet). Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene, 67(1), 31‑40. https://doi.org/10.1016/0378-1119(88)90005-4
• Harper, S. & Speicher, D. W. (2010, 8 octobre). Purification of Proteins Fused to Glutathione S-Transferase. Methods in Molecular Biology, 259‑280. https://doi.org/10.1007/978-1-60761-913-0_14
• Terpe, K. (2003, janvier). Overview of tag protein fusions : from molecular and biochemical fundamentals to commercial systems. Applied Microbiology and Biotechnology, 60(5), 523‑533. https://doi.org/10.1007/s00253-002-1158-6
• Lu, Q., Bauer, J. C. & Greener, A. Using Schizosaccharomyces pombe as a host for expression and purification of eukaryotic proteins. Gene 200, 135–144 (1997).
• Rudert, F. et al. pLEF, a novel vector for expression of glutathione S-transferase fusion proteins in mammalian cells. Gene 169, 281–282 (1996).