Difference between revisions of "Part:BBa K200028"

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<partinfo>BBa_K200028 short</partinfo>
 
<partinfo>BBa_K200028 short</partinfo>
  
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Site directed mutagenesis was carried out to change a serine residue (Ser16) to a glutamine to make the PAH enzyme protease resistant. This will allow safe passage through the stomach and gut in order to carry out function as required by Imperial College's 2009 iGEM project, [http://2009.igem.org/Team:Imperial_College_London <i>The E.ncapsulator</i>].
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The Homo <i>sapiens</i> endogenous enzyme is synthesised by the liver.
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However, as part of the Imperial College iGEM 2009 project, [http://2009.igem.org/Team:Imperial_College_London <i>The E.ncapsulator</i>], the enzyme is released in the intestine of the individual. The presence of proteases in the intestine was therefore a serious problem which would jeopardise the viability of the curative enzyme.
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We thus performed site directed mutagenesis in order to alter a serine residue (Ser16) to a glutamine. This change has previously been correlated with resistance against intestinal proteases<cite>#PRPAH1</cite>.
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In fact, it is thought that phosphorylation of the serine 16 is responsible for the resistance to proteases<cite>#PRPAH2</cite>. This phosphorylated state is mimicked by replacing the serine with a glutamine residue.
  
 
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===Usage and Biology===
 
===Usage and Biology===
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This section is explained in greater detail at the main PAH part page ([https://parts.igem.org/Part:BBa_K200008 BBa_K200008])
  
 
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<partinfo>BBa_K200028 parameters</partinfo>
 
<partinfo>BBa_K200028 parameters</partinfo>
 
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<biblio>
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#PRPAH1 pmid=7887915
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#PRPAH2 pmid=8573072
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</biblio>

Latest revision as of 23:48, 18 October 2009

Protease resistant PAH

The Homo sapiens endogenous enzyme is synthesised by the liver. However, as part of the Imperial College iGEM 2009 project, [http://2009.igem.org/Team:Imperial_College_London The E.ncapsulator], the enzyme is released in the intestine of the individual. The presence of proteases in the intestine was therefore a serious problem which would jeopardise the viability of the curative enzyme. We thus performed site directed mutagenesis in order to alter a serine residue (Ser16) to a glutamine. This change has previously been correlated with resistance against intestinal proteases#PRPAH1. In fact, it is thought that phosphorylation of the serine 16 is responsible for the resistance to proteases#PRPAH2. This phosphorylated state is mimicked by replacing the serine with a glutamine residue.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 287
    Illegal BamHI site found at 814
    Illegal XhoI site found at 524
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


<biblio>

  1. PRPAH1 pmid=7887915
  2. PRPAH2 pmid=8573072

</biblio>