Difference between revisions of "Part:BBa K3487010"

Revision as of 09:29, 20 October 2020

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.

@@ Line 16: / Line 16: @@
 ===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.
+[[File:T--SZPT-CHINA--RS-RalR.png|thumb|center|500px|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===