Team:Mississippi State/Try

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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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Revision as of 18:26, 27 June 2008

Sam i got something.

'''iGEM?'''

Hundreds of undergraduates all over the world spend their summer making Synthetic Biology a reality by participating in the annual International Genetically Engineered Machine competition.

$1.40

Mississippi State University Synthetic Biology

You can write a background of your team here. Give us a background of your team, the members, etc. Or tell us more about something of your choosing.

Msusb logo.JPG


Project Goals

  1. Fuse GFPuv to GhR1 (the gene corresponding with our E3 of choice).
  2. Clone GhR1-GFPuv gene into plasmid.
  3. Sequence to check.
  4. Fuse RFP to ubiquitin.
  5. Clone RFP-ubiquitin gene into separate plasmid.
  6. Sequence to check.
  7. Express proteins in the cell. They will interact with each other and the poly-ubiquitin chain will grow.
  8. Lyse the cells.
  9. Add components for binding.
  10. Run gel.
  11. Analyze gel under UV light.
  • 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.

Alternatives to co-transforming 2 plasmids into the cell:
a) Perform 2 transformations in 1 pET plasmid.
b) Find another plasmid that has a location to clone RFP into.


[http://www.abe.msstate.edu/wiki/msusbcwiki/index.php/Problems
Problems Encountered + Solutions]

  • June 22 - Ubiquitin-J01095 Construction
  • July 6 - Low GhR1 Expression in Protein Gel
  • July 23 - Unsuccessful Amplification of Ubiquitin
  • July 25 - Lack of Protein Separation
  • July 31 - Incomplete Protein Expression
  • August 8 - Restriction Site Within GhR1 Gene
  • August 28 - Incorrect GhR1 Sequence
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