Part:BBa_K2355002
AmilCP with 6x His-tag
This is a modification of the AmilCP part from the Uppsala 2011 team (BBa_K592009). It has a 6x His-tag attached to the C-terminal end of the protein
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Results of experiments
We aimed to improve the amilCP part from the distribution kit by adding a 6x his-tag to the C-terminal end of the chromoprotein. By doing this, it would theoretically be possible to purify both the amilCP part, and composite parts based on the his-tagged amilCP, using affinity chromatography.
The first picture is amilCP with His-tag spun down in LB media.
The second is a picture of amilCP with His-tag after the lysis procedure by sonication.
It is clear that the expression of the blue/purple color is very strong. In conclusion the amilCP has been successfully modified by adding a His-tag without compromising the protein.
To test whether the amilCP with His-tag worked as expected, we purified two solutions of amilCP; one with His-tagged amilCP, and one with just amilCP.
Figure 3: Comparison of how amilCP with or without His-tag is purified. The data used graph is from the samples with the highest amount of sample loading (5 times). The absorbance was measured at 588 nm.
From these results we can conclude that our his-tag purification of amilCP has worked.
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