Difference between revisions of "Part:BBa T2016"
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This is a S-tag head domain designed to be at the N-terminus of a protein. S-tags are a type of affinity tag that can be used to purify and/or measure the quantity of proteins. | This is a S-tag head domain designed to be at the N-terminus of a protein. S-tags are a type of affinity tag that can be used to purify and/or measure the quantity of proteins. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
| + | The protease subtilisin cleaves a single peptide bond in native RNase A. The resulting two fragments are the S-peptide (residues 1-20 of RNase A) and the S-protein (residues 21-124 of RNase A). These two fragments noncovalently associate to form a functional ribonuclease, called RNase S. | ||
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| + | The S-tag system was developed based on the high affinity between the S-peptide and S-protein of RNase S. The S-tag is derived from the first 15 residues of the S-peptide and therefore was originally called S15<cite>Kim93</cite>. It binds with high affinity to the S-protein (or S-fragment) of RNase A. Elution from the S-protein occurs at low pH. An RNase A assay is possible for a quantitative assay of protein expression levels. Colorimetric assays are also available for detection of S-tagged proteins without the use of an antibody. | ||
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Revision as of 13:05, 3 September 2009
S-tag head domain (KETAAAKFERQHMDS)
This is a S-tag head domain designed to be at the N-terminus of a protein. S-tags are a type of affinity tag that can be used to purify and/or measure the quantity of proteins.
Usage and Biology
The protease subtilisin cleaves a single peptide bond in native RNase A. The resulting two fragments are the S-peptide (residues 1-20 of RNase A) and the S-protein (residues 21-124 of RNase A). These two fragments noncovalently associate to form a functional ribonuclease, called RNase S.
The S-tag system was developed based on the high affinity between the S-peptide and S-protein of RNase S. The S-tag is derived from the first 15 residues of the S-peptide and therefore was originally called S15Kim93. It binds with high affinity to the S-protein (or S-fragment) of RNase A. Elution from the S-protein occurs at low pH. An RNase A assay is possible for a quantitative assay of protein expression levels. Colorimetric assays are also available for detection of S-tagged proteins without the use of an antibody.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 16
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 10