Composite

Part:BBa_K3487010

Designed by: Jianhua Zhou   Group: iGEM20_SZPT-CHINA   (2020-10-09)
Revision as of 09:29, 20 October 2020 by Linruixin (Talk | contribs)


A kill switch regulated by lysine concentration

Description

RS is a riboswitch regulated by lysine, which regulates the concentration of lysine to turn on or off downstream gene expression;RalR is a toxic protein that can cleave methylated and unmethylated DNA, leading to cell death.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 12
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 284
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 256

Result

We have constructed a suicide mechanism to ensure that the transgenic bacteria will not be harmful to the environment. The results of the construction of the recombinant plasmid pET28a-RS-RalR are shown in the following.

1% agarose gel image showing the results of PCR and double digestion M,D2000 Plus Marker; Line 1~3, pET28a（+）-RS-RalR double restriction digestion verification; Line 4~6，RS-RalR PCR verification

Reference

Guo Y, Quiroga C, Chen Q, et al. RalR (a DNase) and RalA (a small RNA) form a type I toxin-antitoxin system in Escherichia coli. Nucleic Acids Res. 2014;42(10):6448-6462.
Garst AD, Héroux A, Rambo RP, Batey RT. Crystal structure of the lysine riboswitch regulatory mRNA element. J Biol Chem. 2008;283(33):22347-22351.


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