Difference between revisions of "Part:BBa K200014:Design"

 
(→‎Design Notes)
 
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<partinfo>BBa_K200014 short</partinfo>
 
<partinfo>BBa_K200014 short</partinfo>
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===Design Notes===
 
===Design Notes===
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The cleavable linker facilitiates the synthesis of this pentapeptide as it does not begin with the amino acid methionine.
 
  
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==Cleavable Linker==
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Since polypeptides can only be biologically synthesised if they start with the amino acid methionine, it has been necessary to design a cleavable peptide sequence starting with methionine. This sequence is found upstream of opiorphin and is enzymatically cleaved following incubation with the enzyme enterokinase. Enterokinase is a highly specific serine protease that cleaves after the recognition sequence Aspartic Acid - Aspartic Acid - Aspartic Acid -Aspartic Acid – Lysine.
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==Polyhistidine-tag==
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Six histidine residues are found upstream of the EK recognition sequence. Since histidine is a charged amino acid, it can be used in affinity purification to obtain high grade opiorphin.
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==Stop Codon==
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A double stop codon has been incorporated into the sequence to ensure efficient termination.
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==Codon Choice==
  
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The sequence has been optimised for expression in <i>E.coli</i>.
  
 
===Source===
 
===Source===

Latest revision as of 15:43, 18 October 2009

Opiorphin + EK cleavage site + His Tag


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
    COMPATIBLE WITH RFC[1000]


Design Notes

Cleavable Linker

Since polypeptides can only be biologically synthesised if they start with the amino acid methionine, it has been necessary to design a cleavable peptide sequence starting with methionine. This sequence is found upstream of opiorphin and is enzymatically cleaved following incubation with the enzyme enterokinase. Enterokinase is a highly specific serine protease that cleaves after the recognition sequence Aspartic Acid - Aspartic Acid - Aspartic Acid -Aspartic Acid – Lysine.

Polyhistidine-tag

Six histidine residues are found upstream of the EK recognition sequence. Since histidine is a charged amino acid, it can be used in affinity purification to obtain high grade opiorphin.


Stop Codon

A double stop codon has been incorporated into the sequence to ensure efficient termination.


Codon Choice

The sequence has been optimised for expression in E.coli.

Source

Synthetic sequence.

References