Difference between revisions of "Part:BBa T2015"

Revision as of 12:54, 3 September 2009

S-tag tail domain (KETAAAKFERQHMDS)

This is a S-tag tail domain designed to be at the C-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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NotI site found at 10
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 4

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 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 of 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.
+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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