Difference between revisions of "Part:BBa K4656011"

Latest revision as of 11:50, 22 September 2023

pRha-tetR-tetO-mazF

L-rhamnose-inducible promoter is capable of high-level recombinant protein expression in the presence of L-rhamnose，it is also tightly regulated in the absence of L-rhamnose by the addition of D-glucose, which has been well characterized by Paris_Bettencour in 2012. Tetracycline repressor (TetR) is an effector-regulated DNA-binding protein that binds tightly to its palindromic tetO operator DNA in the absence of an effector, tet or its analogs, thereby blocking the transcription of any downstream genes. TetO, placed downstream of TetR, as mentioned before, can be repressed by TetR. Escherichia coli MazF is a toxin protein that cleaves RNA at ACA sequences, thus causing growth inhibition and block protein synthesis[1].

Usage and Biology

In our circuit, the L-rhamnose-inducible promoter can be activated by L-rhamnose addition, leading to elevated expression of TetR, which will bind to tetO operator tightly and subsequently inhibit the downstream MazF expression, enabling bacteria to survive.

Experimental results

Reference

[1] Zhang, Y., Zhang, J., Hoeflich, K. P., Ikura, M., Qing, G., & Inouye, M. (2003). MazF cleaves cellular mRNAs specifically at ACA to block protein synthesis in Escherichia coli. Molecular cell, 12(4), 913–923. https://doi.org/10.1016/s1097-2765(03)00402-7

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 2: / Line 2: @@
 __NOTOC__
 <partinfo>BBa_K4656011 short</partinfo>
+<html><BR><BR><center><img style="display: block;-webkit-user-select: none;margin: auto;cursor: zoom-in;background-color: hsl(0,0%,90%);transition: background-color 300ms;" src="https://static.igem.wiki/teams/4656/wiki/part-prha-tetr.png" width="600" height="150"></center></html>
-L-rhamnose-inducible promoter is capable of high-level recombinant protein expression in the presence of L-rhamnose， it is also tightly regulated in the absence of L-rhamnose by the addition of D-glucose, which has been well characterized by Paris_Bettencour in 2012. Tetracycline repressor (TetR) is an effector-regulated DNA-binding protein that binds tightly to its palindromic tetO operator DNA in the absence of an effector, tet or its analogs, thereby blocking the transcription of any downstream genes. TetO, placed downstream of TetR, as mentioned before, can be repressed by TetR. Escherichia coli MazF is a toxin protein that cleaves RNA at ACA sequences, thus causing growth inhibition and block protein synthesis[1].
+L-rhamnose-inducible promoter is capable of high-level recombinant protein expression in the presence of L-rhamnose，it is also tightly regulated in the absence of L-rhamnose by the addition of D-glucose, which has been well characterized by Paris_Bettencour in 2012. Tetracycline repressor (TetR) is an effector-regulated DNA-binding protein that binds tightly to its palindromic tetO operator DNA in the absence of an effector, tet or its analogs, thereby blocking the transcription of any downstream genes. TetO, placed downstream of TetR, as mentioned before, can be repressed by TetR. Escherichia coli MazF is a toxin protein that cleaves RNA at ACA sequences, thus causing growth inhibition and block protein synthesis[1].
 ===Usage and Biology===
 In our circuit, the L-rhamnose-inducible promoter can be activated by L-rhamnose addition, leading to elevated expression of TetR, which will bind to tetO operator tightly and subsequently inhibit the downstream MazF expression, enabling bacteria to survive.