Difference between revisions of "Part:BBa K2552007"
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | This part is used to regulate azurin(14kDa) expression by Pyear promoter([https://parts.igem.org/Part:BBa_K216005]). | + | This part is used to regulate azurin(14kDa) expression by Pyear promoter([https://parts.igem.org/Part:BBa_K216005 BBa_K216005]). |
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The Pyear is the promoter of the Escherichia coli yeaR/yoaG operon (see Lin, H.-Y., Bledsoe, P.J., and Stewart, V. 2007. Activation of yeaR-yoaG operon transcription by the nitrate-responsive regulator NarL is independent of oxygen-responsive regulator Fnr in Escherichia coli K-12. J. Bacteriol. 189, 7539-7548). | The Pyear is the promoter of the Escherichia coli yeaR/yoaG operon (see Lin, H.-Y., Bledsoe, P.J., and Stewart, V. 2007. Activation of yeaR-yoaG operon transcription by the nitrate-responsive regulator NarL is independent of oxygen-responsive regulator Fnr in Escherichia coli K-12. J. Bacteriol. 189, 7539-7548). | ||
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We want to regulate the expression of azurin protein only with the present of nitric oxide so we combine the Pyear promoter with the azurin protein. We also add the His tag (6 Histidine amino acids) at the end of azurin protein in order to exam the expression of it. | We want to regulate the expression of azurin protein only with the present of nitric oxide so we combine the Pyear promoter with the azurin protein. We also add the His tag (6 Histidine amino acids) at the end of azurin protein in order to exam the expression of it. | ||
| − | We insert the Pyear promoter and azurin protein into a regulative vector pCDFDuet1. After inducing with sedium nitrate, we extract all the protein in our strain and exam it by Western Blotting method. From the Fig1, we demonstrate that azurin protein successfully express with the His tag. | + | We insert the Pyear promoter and azurin protein into a regulative vector pCDFDuet1. |
| + | After inducing with sedium nitrate, we extract all the protein in our strain and exam it by Western Blotting method. From the Fig1, we demonstrate that azurin protein successfully express with the His tag. | ||
[[Image:001wb.jpeg|centre|500px]] | [[Image:001wb.jpeg|centre|500px]] | ||
Latest revision as of 13:54, 17 October 2018
Pyear-azurin
This part is used to regulate azurin expression by Pyear promoter.
Usage and Biology
This part is used to regulate azurin(14kDa) expression by Pyear promoter(BBa_K216005).
The Pyear is the promoter of the Escherichia coli yeaR/yoaG operon (see Lin, H.-Y., Bledsoe, P.J., and Stewart, V. 2007. Activation of yeaR-yoaG operon transcription by the nitrate-responsive regulator NarL is independent of oxygen-responsive regulator Fnr in Escherichia coli K-12. J. Bacteriol. 189, 7539-7548).
We want to regulate the expression of azurin protein only with the present of nitric oxide so we combine the Pyear promoter with the azurin protein. We also add the His tag (6 Histidine amino acids) at the end of azurin protein in order to exam the expression of it. We insert the Pyear promoter and azurin protein into a regulative vector pCDFDuet1. After inducing with sedium nitrate, we extract all the protein in our strain and exam it by Western Blotting method. From the Fig1, we demonstrate that azurin protein successfully express with the His tag.
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Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]