Part:BBa_K3143005
Phd3TAG
Phd3TAG is an ncAA-dependent antitoxin in which 3 TAGs are inseted just after the start codon of Phd. See more about Phd BBa_K3143002, Phd1TAG BBa_K3143003 and Phd2TAG BBa_K3143004.
Characterization
In order to build a Toxin-Antitoxin system which is dependent on ncAA, see why we want to build this system. First we tried to confirm that insertion of how many TAG codons could completely inhibit gene expression. We inserted 1-3 TAG codons just after the start codon, and then measured the fluorescence value of Phd-GS-sfGFP. The results showed the expression of the gene was well terminated in three designs, and the insertion of two TAG codons was the best.
Figure 1: Characterization of Phd-TAG-GS-sfGFP system
Next we attempted to introduce the ncAA system into the Phd-GS-sfGFP expression system with two TAG codons. When the ncAA system is expressed, we can see that the system that originally had no fluorescence produced a strong fluorescent signal. This shows that we have successfully established an Anticoxin (Phd) system that relies on ncAA. When ncAA is present in the system, Phd can be successfully expressed, and when ncAA is not present in the system, the expression of Phd is completely terminated.
Figure 2: Characterization of Phd(2TAG)-Doc system
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]
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