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===
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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.
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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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The S-tag can be used at the N- or C-terminus or internal to a protein of interest.
  
 
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Latest revision as of 13:06, 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.

The S-tag can be used at the N- or C-terminus or internal to a protein of interest.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 16
  • 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 10