Difference between revisions of "Part:BBa K3487007"

Latest revision as of 14:56, 27 October 2020

Lysine riboswitch

Description

Riboswitch is a regulatory part of an mRNA molecule that can specifically bind a metabolite and regulate gene expression. Till now, more than 20 riboswitches have been discovered. Among them, only two amino acids riboswitches were found: a lysine riboswitch and a glycine riboswitch. Several previous studies examined the functionality and manipulation of the B. subtilis lysine riboswitch. A lysine riboswitch was also found in E. coli, which revealed in fact the mechanism of lysine feedback repression of aspartkinase III in E. coli. The following is the control principle diagram

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

2020 SZPT-CHINA

Result

In our project, we set up a suicide mechanism regulated by this lysine riboswitch part. When the bacteria colonize the oral cavity, the riboswitch controlled by lysine is turned off. When the bacteria escape the oral environment, the riboswitch opens and expresses the toxic protein RalR, which is a non-specific endonuclease. In order to determine at what concentration of lysine this switch can be turned on. We connected sfGFP to lysine riboswitch and then tested the fluorescent expression in different concentrations of lysine for 24 hours. The result is as follows.

References

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 4: / Line 4: @@
 ===Description===
-Riboswitches are metabolite-sensitive elements found in mRNAs that control gene expression through a regulatory secondary structural switch.RS is a Lysine riboswitch that Regulated by lysine.
+Riboswitch is a regulatory part of an mRNA molecule that can specifically bind a metabolite and regulate gene expression. Till now, more than 20 riboswitches have been discovered. Among them, only two amino acids riboswitches were found: a lysine riboswitch and a glycine riboswitch. Several previous studies examined the functionality and manipulation of the B. subtilis lysine riboswitch. A lysine riboswitch was also found in <i>E. coli</i>, which revealed in fact the mechanism of lysine feedback repression of aspartkinase III in <i>E. coli</i>.
 The following is the control principle diagram
 [[File:T--SZPT-CHINA--RS.png|500px|center|]]
@@ Line 16: / Line 16: @@
 ==2020 SZPT-CHINA==
 ===Result===
-We put the lysine riboswitch connected sfGFP engineered bacteria in different concentrations of lysine for 24 hours, and then tested the fluorescent expression of the bacteria. The results are shown in the following.
+In our project, we set up a suicide mechanism regulated by this lysine riboswitch part. When the bacteria colonize the oral cavity, the riboswitch controlled by lysine is turned off. When the bacteria escape the oral environment, the riboswitch opens and expresses the toxic protein RalR, which is a non-specific endonuclease.
-[[File:T--SZPT-CHINA--RS2.png|500px|center|]]
+In order to determine at what concentration of lysine this switch can be turned on. We connected sfGFP to lysine riboswitch and then tested the fluorescent expression in different concentrations of lysine for 24 hours. The result is as follows.
-The results show that the bacteria are in the lysine at a concentration of 30μmol/mL. Fluorescence expression was significantly inhibited, indicating that the lysine riboswitch can work normally.
+[[File:T--SZPT-CHINA--RS2.png|700px|center]]
 ===References===