Part:BBa_K2669000:Experience

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Applications of BBa_K2669000

After transforming our cells with a low copy amplicilin plasmid containing this composite part, cell lysis and affinity chromotography were used to extract UnaG from our cells. Please note: The exact procedure can be found at the end of our UnaG wiki page. Conducting "bilirubin tests" (the addition of a small amount of bilirubin dissolved in chloroform to samples) allowed us to see if UnaG was present in our samples, since as mentioned earlier UnaG fluoresces in the presence of bilirubin.

Figure 1: Bilirubin test before/after affinity chromatography. Going from right to left the samples are:

• Lysed sample of the “bad” part before AC
• Lysed sample of the “good” part before AC
• "Bad" part after AC
• "Good" part after AC

Figure 2: Comparison of blank tube to successful extraction/previous iGEM part. The tubes reading from left to right are as followed:

• Blank tube with AC elution buffer/bilirubin
• Tube with bilirubin + original iGEM UnaG part
• Our extracted modified UnaG with a moved start codon, as can be seen in Figure 1

A good degree of fluorescence can be seen in the last tube compared to the other two, which clearly contain none of our protein of interest.

User Reviews

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