Difference between revisions of "Part:BBa K3292004"
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This part is an improvement of the BBa_I746916 which allows you to choose which is the better option for your protein either to purify it with an affinity column or just to take it from the medium. | This part is an improvement of the BBa_I746916 which allows you to choose which is the better option for your protein either to purify it with an affinity column or just to take it from the medium. | ||
− | + | ||
− | === | + | ===Improvement of BBa_I746916 by iGEM TecMonterrey=== |
+ | |||
+ | The BioBrick BBa_I746916 corresponds to the coding sequence for the sfGFP. According to Zhang et al. (2019), this protein presents an auto-secretory activity, | ||
+ | and can also secrete other proteins when linked to them. Based on this, we decided to improve this part by adding a 6x his-tag to facilitate other teams the recovery | ||
+ | of proteins of interest, and compare it with other traditional purification methods. | ||
+ | |||
+ | To charactherize this improvement we first performed a fluorescence assay in order to observe the production of this protein by induction with IPTG | ||
+ | at different concentrations. In addition we performed an SDS-PAGE to observe our expressed protein under the same induction conditions by different concentrations of IPTG. | ||
+ | Finally, we performed a Bradford essay to quantify our protein. | ||
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Revision as of 02:37, 22 October 2019
6His-tagged Super Folding GFP with terminator
This part is an improvement of the BBa_I746916 which allows you to choose which is the better option for your protein either to purify it with an affinity column or just to take it from the medium.
Improvement of BBa_I746916 by iGEM TecMonterrey
The BioBrick BBa_I746916 corresponds to the coding sequence for the sfGFP. According to Zhang et al. (2019), this protein presents an auto-secretory activity, and can also secrete other proteins when linked to them. Based on this, we decided to improve this part by adding a 6x his-tag to facilitate other teams the recovery of proteins of interest, and compare it with other traditional purification methods.
To charactherize this improvement we first performed a fluorescence assay in order to observe the production of this protein by induction with IPTG at different concentrations. In addition we performed an SDS-PAGE to observe our expressed protein under the same induction conditions by different concentrations of IPTG. Finally, we performed a Bradford essay to quantify our protein.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 13