Part:BBa_K2958000
TEV with altered amino acid (ENLYFQS instead of ENLYFQG)
Site-specific TEV purification tag with the amino acid sequence ENLYFQS. This TEV protease cleaves between the Glutamine and Serine residues.
Description:
The TEV tag is a cleavage site that allows for the site-specific cleavage of maltose-binding protein (MBP) or poly-histidine purification tags from fusion proteins. The most commonly used recognition site for the TEV tag is ENLYFQG, but this TEV tag was modified to have greater efficiency with the optimal amino acid sequence ENLYFQS based on the journal article, “Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems” by Terpe, K (2002). This TEV tag was incorporated in our insulin construct designs (BBa_K2958009, BBa_K2958010, BBa_K2958011, and BBa_K2958012) in order to cleave at the site of our [6GGs + His tag + 6GGS] purification tag and aid in insulin purification.
References:
Terpe, K. (2003). Overview of tag protein fusions: From molecular and biochemical fundamentals to commercial systems. Applied Microbiology and Biotechnology, 60(5), 523- 33. doi:10.1007/s00253-002-1158-6
Waugh D. S. (2011). An overview of enzymatic reagents for the removal of affinity tags. Protein expression and purification, 80(2), 283–293. doi:10.1016/j.pep.2011.08.005
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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