Difference between revisions of "Part:BBa K2107000:Experience"
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Finally, to obtain more definitive conformation that BBa_k2107000 is being produced and correctly folded we purchased PDGF-B specific antibodies from Santa Cruz Biolabs and ran a western blot. To our delight we were able to visualized a band of protein at the correct location when compared to an industry standard western blot of PDGF-B using the same antibody. This indicates that BBa_K2107000 is being expressed and properly folded in the periplasm. | Finally, to obtain more definitive conformation that BBa_k2107000 is being produced and correctly folded we purchased PDGF-B specific antibodies from Santa Cruz Biolabs and ran a western blot. To our delight we were able to visualized a band of protein at the correct location when compared to an industry standard western blot of PDGF-B using the same antibody. This indicates that BBa_K2107000 is being expressed and properly folded in the periplasm. | ||
− | </br></br>       <img src="https://static.igem.org/mediawiki/parts/a/a5/Lubbock_ttu_western_blot_messy.png" width="" height""></img>                <img src="https://static.igem.org/mediawiki/parts/4/47/Lubbock_ttu_western_blot_clean.png" width="" height""></img></br><center><b>Fig 3.</b> Western Blot example of PDGF-B from Santa Cruz biotechnology.    <b>Fig 4.</b> Western blot of recombinant PDGF-B Iso 1 purified from E. coli </center></br></br> | + | </br></br>       <img src="https://static.igem.org/mediawiki/parts/a/a5/Lubbock_ttu_western_blot_messy.png" width="" height""></img>                 <img src="https://static.igem.org/mediawiki/parts/4/47/Lubbock_ttu_western_blot_clean.png" width="" height""></img></br><center><b>Fig 3.</b> Western Blot example of PDGF-B from Santa Cruz biotechnology.    <b>Fig 4.</b> Western blot of recombinant PDGF-B Iso 1 purified from E. coli </center></br></br> |
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Revision as of 23:46, 30 October 2016
Validation and proof of concept of BBa_K2107000 1. Construct validation: After cloning and transformation we did growth curves using this part. Control groups consisted of the cell chassis alone and cell chassis with inducer. Test groups consisted of the transformed cell chassis with and without inducer. We saw that the cell transformed cell chassis exhibited a slight fitness cost indicating acceptance of BBa_K2107000. While the transformed cell chassis with inducer exhibited a large fitness cost indicating that the construct is being expressed.
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