Difference between revisions of "Part:BBa K4698007"
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<p>While reviewing CRISPR-related materials, we discovered a particularly interesting evolutionary system- phage-assisted continuous evolution (PACE)[1,2], which can enable continuous directed evolution of gene-coding molecules that can be associated with protein production in <i>E. coli</i>. Then, we found a protein degradation tag-degron[3], that enables rapid hydrolysis of proteins with little functionality, which allows degron to be used in a number of protein co-expression screening systems.</p> | <p>While reviewing CRISPR-related materials, we discovered a particularly interesting evolutionary system- phage-assisted continuous evolution (PACE)[1,2], which can enable continuous directed evolution of gene-coding molecules that can be associated with protein production in <i>E. coli</i>. Then, we found a protein degradation tag-degron[3], that enables rapid hydrolysis of proteins with little functionality, which allows degron to be used in a number of protein co-expression screening systems.</p> | ||
− | <p>We | + | <p>We expressed GFP in fusion with degron and tested the degradation efficiency of degron (Fig. 1). The results showed that the GFP-degron fusion protein was almost non-fluorescent, much less than GFP. this demonstrated the extremely strong degradation activity of degron.</p> |
<img src="https://static.igem.wiki/teams/4698/wiki/improved-part-gfp.png" class= "center" style="width:500px"> | <img src="https://static.igem.wiki/teams/4698/wiki/improved-part-gfp.png" class= "center" style="width:500px"> | ||
<p>Figure 1, Testing the protein degradation efficiency of degron. blank, LB medium. NT, <i>E.coli</i> without GFP expression plasmid, <i>n</i>=3.</p> | <p>Figure 1, Testing the protein degradation efficiency of degron. blank, LB medium. NT, <i>E.coli</i> without GFP expression plasmid, <i>n</i>=3.</p> |
Latest revision as of 15:26, 11 October 2023
GFP with a protein degradation tag attached
A protein degradation tag (degron) is attached to the C-terminus of GFP to promote protein degradation.
Usage and Biology
While reviewing CRISPR-related materials, we discovered a particularly interesting evolutionary system- phage-assisted continuous evolution (PACE)[1,2], which can enable continuous directed evolution of gene-coding molecules that can be associated with protein production in E. coli. Then, we found a protein degradation tag-degron[3], that enables rapid hydrolysis of proteins with little functionality, which allows degron to be used in a number of protein co-expression screening systems.
We expressed GFP in fusion with degron and tested the degradation efficiency of degron (Fig. 1). The results showed that the GFP-degron fusion protein was almost non-fluorescent, much less than GFP. this demonstrated the extremely strong degradation activity of degron.
![](https://static.igem.wiki/teams/4698/wiki/improved-part-gfp.png)
Figure 1, Testing the protein degradation efficiency of degron. blank, LB medium. NT, E.coli without GFP expression plasmid, n=3.
Refrence
1. Esvelt, K. M., Carlson, J. C. & Liu, D. R. A system for the continuous directed evolution of biomolecules. Nature 472, 499-503 (2011). 2. Miller, S. M., Wang, T. & Liu, D. R. Phage-assisted continuous and non-continuous evolution. Nature Protocols 15, 4101-4127 (2020). 3. Karzai, A. W., Roche, E. D. & Sauer, R. T. The SsrA-SmpB system for protein tagging, directed degradation and ribosome rescue. Nat Struct Biol 7, 449-455 (2000).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]