Difference between revisions of "Part:BBa K1800000:Experience"
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The 2015 GSU iGEM team was able to successfully express and characterize the mambalgin protein in E. coli. After inducing with IPTG overnight, the cell cultures were centrifuged and disrupted with a French press. Mambalgin was isolated using affinity chromatography, utilizing the 6x His tag in the construct. A SDS-PAGE was done, followed by a Ponceau S. and coomassie stain to visualize the proteins. A band indicating a protein of ~9 kda – the size of the mambalgin for E. coli construct – was seen in the eluate fraction | The 2015 GSU iGEM team was able to successfully express and characterize the mambalgin protein in E. coli. After inducing with IPTG overnight, the cell cultures were centrifuged and disrupted with a French press. Mambalgin was isolated using affinity chromatography, utilizing the 6x His tag in the construct. A SDS-PAGE was done, followed by a Ponceau S. and coomassie stain to visualize the proteins. A band indicating a protein of ~9 kda – the size of the mambalgin for E. coli construct – was seen in the eluate fraction | ||
| + | https://parts.igem.org/File:Gsuigemgel.png | ||
https://static.igem.org/mediawiki/2015/d/dd/MAMBA.E.coli_Western_blot.THISONE.jpg | https://static.igem.org/mediawiki/2015/d/dd/MAMBA.E.coli_Western_blot.THISONE.jpg | ||
Revision as of 23:23, 26 September 2015
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The 2015 GSU iGEM team was able to successfully express and characterize the mambalgin protein in E. coli. After inducing with IPTG overnight, the cell cultures were centrifuged and disrupted with a French press. Mambalgin was isolated using affinity chromatography, utilizing the 6x His tag in the construct. A SDS-PAGE was done, followed by a Ponceau S. and coomassie stain to visualize the proteins. A band indicating a protein of ~9 kda – the size of the mambalgin for E. coli construct – was seen in the eluate fraction
https://parts.igem.org/File:Gsuigemgel.png
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