Part:BBa_K526001
ptrc*-Dpn
ptrc* is a modified form of ptrc promoter with the -35 and -10 regions being present in the reverse order and the promoter is flanked by fimE recognition sequence. FimE is an invertase capable of inverting any DNA sequence flanked between its recognition seqence. This fim inversion system would help avoid leaky expression. We expressed a new lysis device, DpnI under the control of fim inversion promoter. DpnI is a restriction enzyme capable of chopping up all methylated DNAs. We propose DpnI as a novel lysis device. DpnC and DpnD are obtained from Streptococcus pneumoniae R6 strain.
Significance of this Device
1. There are lot of lysis devices engineered before such as Holin mediated lysis device, lysozyme mediated lysis device and kill genes mediated self killing devices.
2. All these devices cause cell lysis. None of these devices had considered the eradication of DNA, the more stable genetic material.
3.DNA is supposed to resist all harsh environment and stay for long with out being degraded.
4. These stable DNA could be taken up by other microbes in the environment through a process called Horizontal Gene Transfer (HGT).
5. HGT is one big fear that causes synthetic biology to lag behind. One would not forget the threats imposed by GMOs on Monarch butterflies and Gulf bacterium.
6. Our self-lysis device might be superior to other lysis devices as it helps in chopping up the DNA completely.
7. This would ensure that there is no HGT and would help in more direct application of the synthetic microbes to the environment.
8. Dpn mediated cell death, if optimized, would help in designing synthetic microbes that could be used in the environment without any fear.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1542
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 820
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