Part:BBa_K5291037:Design
pAB-hok/sok
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 146
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 16
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
We replace the promoter of sok by PopdH, a promoter able to sense the concentration of citrate, and replace the promoter of hok/mok by PcW, a common weak constitutive promoter in Pseudomonas aeruginosa. When the bacteria are in the mangrovem, the relatively high concentration of citrate is able to activate PopdH and sok can be expressed to antagonize the effect of hok/mok due to the weak function of PcW. When the bacteria are out of the mangroves, PopdH cannot come into effect and the expression of sok is terminated, which enables hok to kill the cells.
When the plasmid with hok/sok system is lost, both of the genes cannot be expressed. However, the half-life of the hok mRNA is longer than the one of the sok antisense RNA, which leads to the result that the hok mRNA still exists when all the sok antisense RNA is degraded. The rest hok mRNA can encode toxin to kill the cell, achieving the function of plasmid anti-loss.
Source
PopdH, PcW: Pseudomonas aeruginosa PAO1
sok, hok/mok, BiTerm: Escherichia coli
References
[1]Puskas, L.G., et al., A periplasmic, alpha-type carbonic anhydrase from Rhodopseudomonas palustris is essential for bicarbonate uptake. Microbiology (Reading), 2000. 146 ( Pt 11): p. 2957-2966.
[2]Thorbecke, R., et al., The gene dosage effect of carbonic anhydrase on the biosynthesis of poly ( 3-hydroxybutyrate ) under autotrophic and mixotrophic culture conditions. POLYMER JOURNAL, 2021. 53(1): p. 209-213.
[3]Chen YJ, Liu P, Nielsen AA, Brophy JA, Clancy K, Peterson T, Voigt CA. Characterization of 582 natural and synthetic terminators and quantification of their design constraints. Nat Methods. 2013 Jul;10(7):659-64. doi: 10.1038/nmeth.2515. Epub 2013 Jun 2. PMID: 23727987.