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Team:Slovenia/PROJECT/proof/studies/prod
From 2010.igem.org
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| + | <p><strong><span style="font-size: 16px; line-height: 25px;">Cloning scheme</span></strong></p> | ||
<p> </p> | <p> </p> | ||
| + | Before any <em>in vitro</em> characterisation, DNA-binding proteins had to be obtained. Zinc fingers were firstly tri-point ligated with a particular split GFP and T7 promoter and T7 terminator sequence were added afterwards. T7 promoter enables high production of proteins in<em> E. coli </em>BL21(DE3)pLysS production strain. Zinc fingers were then purified using His tags and used in subsequent experiments. Parts used for production were: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K323001 BBa_323001], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K323015 BBa_323015], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K323069 BBa_323069], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K323058 BBa_323058], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K323004 BBa_323004], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K323070 BBa_323070]. | ||
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| + | [[Image:slo_dna_scafold2.jpg|thumb|center|700px]] | ||
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Revision as of 12:40, 27 October 2010
Cloning scheme
Before any in vitro characterisation, DNA-binding proteins had to be obtained. Zinc fingers were firstly tri-point ligated with a particular split GFP and T7 promoter and T7 terminator sequence were added afterwards. T7 promoter enables high production of proteins in E. coli BL21(DE3)pLysS production strain. Zinc fingers were then purified using His tags and used in subsequent experiments. Parts used for production were: BBa_323001, BBa_323015, BBa_323069, BBa_323058, BBa_323004, BBa_323070.
