Difference between revisions of "Part:BBa K1456006:Experience"
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<li>HEK 293T cells have been cotransfected with 1 µg, 2 µg, 3µg ve 4 µg Tet off – pTRE Aprotinin. The cells collected and protein isolation was performed to measure the serine protease inhibition. According to the positive control test, these results Show dose-dependent reduction and the production of aprotinin has increased according to the amount of cells successfully transfected. | <li>HEK 293T cells have been cotransfected with 1 µg, 2 µg, 3µg ve 4 µg Tet off – pTRE Aprotinin. The cells collected and protein isolation was performed to measure the serine protease inhibition. According to the positive control test, these results Show dose-dependent reduction and the production of aprotinin has increased according to the amount of cells successfully transfected. | ||
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+ | </br><h2>Combination of DsbA Signal Peptide with Aprotinin to Enhance Protein Purification (Lubbock_TTU iGEM 2016)</h2> | ||
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+ | The Aprotinin BioBrick by the ATOMS-Turkiye 2014 iGEM team was enhanced for better protein purification by using a DsbA signal tag in combination with the Aprotinin BioBrick. DsbA signal peptide carries Aprotinin to the periplasm where proper protein folding is enhanced. A protein extraction method known as osmotic shock creates pores in the outer membrane releasing the Aprotinin and other proteins in the periplasm into the supernatant. There should be fewer proteins in the solution compared with using a cell lysis method that releases all proteins within the cell into the solution. This results in cleaner protein purification of Aprotinin, while maintaining functionality. | ||
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+ | <center><br><img src="https://static.igem.org/mediawiki/parts/5/5d/Lubbock_TTU_aprotinin_growth_curve.png" height="" width="50%"></br><b>Fig 1.</b> In the above figure we see that IPTG induced origami cells containing the Aprotinin expression vector </br>exhibit the greatest fitness cost to proliferation rate, indicating that significant resources are being allocated to protein production. n=3</center> | ||
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+ | </br><h2>Simplification and E. coli optimization(Alma iGEM 2024)</h2> | ||
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+ | The Aprotinin BioBrick by the ATOMS-Turkiye 2014 was redesigned to be a more simplified part. This includes removing the DsbA signal peptide along with the histidine signal at the end of the protein. This fix removes the reading frame shift the part had. A codon optimization for E. coli was performed to make the final part <partinfo>BBa_K5324000</partinfo> . | ||
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+ | </html> | ||
===Applications of BBa_K1456006=== | ===Applications of BBa_K1456006=== |
Latest revision as of 00:03, 26 September 2024
Aprotinin Assay
Chacarterisation
Combination of DsbA Signal Peptide with Aprotinin to Enhance Protein Purification (Lubbock_TTU iGEM 2016)
The Aprotinin BioBrick by the ATOMS-Turkiye 2014 iGEM team was enhanced for better protein purification by using a DsbA signal tag in combination with the Aprotinin BioBrick. DsbA signal peptide carries Aprotinin to the periplasm where proper protein folding is enhanced. A protein extraction method known as osmotic shock creates pores in the outer membrane releasing the Aprotinin and other proteins in the periplasm into the supernatant. There should be fewer proteins in the solution compared with using a cell lysis method that releases all proteins within the cell into the solution. This results in cleaner protein purification of Aprotinin, while maintaining functionality.Fig 1. In the above figure we see that IPTG induced origami cells containing the Aprotinin expression vector exhibit the greatest fitness cost to proliferation rate, indicating that significant resources are being allocated to protein production. n=3
Simplification and E. coli optimization(Alma iGEM 2024)
The Aprotinin BioBrick by the ATOMS-Turkiye 2014 was redesigned to be a more simplified part. This includes removing the DsbA signal peptide along with the histidine signal at the end of the protein. This fix removes the reading frame shift the part had. A codon optimization for E. coli was performed to make the final partApplications of BBa_K1456006
User Reviews
UNIQ0814e5cad114cb53-partinfo-00000001-QINU
UNIQ0814e5cad114cb53-partinfo-00000002-QINU