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| | *[http://www.uni-bonn.de/~manfear/html2wiki-tables.php HTML to WIKI Converter] | | *[http://www.uni-bonn.de/~manfear/html2wiki-tables.php HTML to WIKI Converter] |
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| - | <h3>Project Goals</h3>
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| - | # Fuse GFPuv to GhR1 (the gene corresponding with our E3 of choice).<br>
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| - | # Clone GhR1-GFPuv gene into plasmid.<br>
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| - | # Sequence to check.<br>
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| - | # Fuse RFP to ubiquitin.<br>
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| - | # Clone RFP-ubiquitin gene into separate plasmid.<br>
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| - | # Sequence to check.<br>
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| - | # Express proteins in the cell. They will interact with each other and the poly-ubiquitin chain will grow.<br>
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| - | # Lyse the cells.<br>
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| - | # Add components for binding.<br>
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| - | # Run gel.<br>
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| - | # Analyze gel under UV light.<br>
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| - | * If the protein band is tagged with GFPuv, it will be visible. The expected outcome of expression of both GFPuv and RFP is amber colored light.
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| - | Alternatives to co-transforming 2 plasmids into the cell:<br>
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| - | a) Perform 2 transformations in 1 pET plasmid.<br>
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| - | b) Find another plasmid that has a location to clone RFP into.
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| - | [http://www.abe.msstate.edu/wiki/msusbcwiki/index.php/Problems<br>'''Problems Encountered + Solutions''']
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| - | * June 22 - Ubiquitin-J01095 Construction
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| - | * July 6 - Low GhR1 Expression in Protein Gel
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| - | * July 23 - Unsuccessful Amplification of Ubiquitin
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| - | * July 25 - Lack of Protein Separation
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| - | * July 31 - Incomplete Protein Expression
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| - | * August 8 - Restriction Site Within GhR1 Gene
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| - | * August 28 - Incorrect GhR1 Sequence
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