Difference between revisions of "Part:BBa K200028"
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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. | 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. | ||
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>. | 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>. | ||
| − | + | 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=== | ||
This section is explained in greater detail at the main PAH part page ([https://parts.igem.org/Part:BBa_K200008 BBa_K200008]) | 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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<biblio> | <biblio> | ||
#PRPAH1 pmid=7887915 | #PRPAH1 pmid=7887915 | ||
| + | #PRPAH2 pmid=8573072 | ||
</biblio> | </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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 287
Illegal BamHI site found at 814
Illegal XhoI site found at 524 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
<biblio>
- PRPAH1 pmid=7887915
- PRPAH2 pmid=8573072
</biblio>