Difference between revisions of "Part:BBa K5322002"

Revision as of 11:14, 29 September 2024

_NOTOC__ Constitutive Mfp3 Expression System

Nitric-oxide-inducible lysis module

Usage and Biology

Plasm11:14, 29 September 2024 (UTC)~

Construction of the plasmid

In o3119-``````````

Figure 2-1 Plasmid pET29(a)-pJ23119-SoxR-T-pSoxS-RBS-Mfp3-T7

Figure 2-2 Colony PCR gel electrophoresis of plasmid pET29(a)-pJ23119-SoxR-T-pSoxS-RBS-PhiX174E-T7(1320bp)

Figure 2-3 plasmid pET29(a)-pJ23119-SoxR-T-pSoxS-RBS-PhiX174E-T7 sequencing result

Protein Expression Validation

We verified the performance of the lysis module through dry experiments and plan to complete wet experimental validation of its lytic function in the future. Through literature review and numerical modeling validation, we found that after inducing the expression of antimicrobial peptides and lysis proteins with NO for twenty minutes, the engineered strain will be lysed by the lysis protein and release the antimicrobial peptides. Mathematical modeling confirmed that at this time, the concentration of antimicrobial peptides is sufficient to reach an effective inhibitory concentration.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 572
21
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 1009
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 1: / Line 1: @@
+_NOTOC__
+<partinfo>BBa_K5322000 short</partinfo>
-__NOTOC__
+Nitric-oxide-inducible lysis module
-<partinfo>BBa_K5322002 short</partinfo>
+<html>
+</p>
+</html>
+__TOC__
-<!-- Add more about the biology of this part here
+==Usage and Biology==
-===Usage and Biology===
+<p>
+Plasm11:14, 29 September 2024 (UTC)~
+</P>
-<!-- -->
+==Construction of the plasmid==
-<span class='h3bb'>Sequence and Features</span>
+<html>
-<partinfo>BBa_K5322002 SequenceAndFeatures</partinfo>
+<p>
+In o3119-``````````
+</p>
+<style>
+  .center-img {
+    text-align:center;
+  }
+</style>
+<div class="center-img">
+<img src="https://static.igem.wiki/teams/5322/wet-lab/20-pet29a-j23119-rbs-mfp3-t7.png" alt="pET29a-J23119-RBS-Mfp3-T7" width="300">
+<p align="center"><b>Figure 2-1</b>  Plasmid pET29(a)-p<i>J23119-SoxR</i>-T-p<i>SoxS</i>-RBS-<i>Mfp3</i>-T7</p>
+</div>
+<div class="center-img">
+<img src="https://static.igem.wiki/teams/5101/partpage/phix174e-gel.png" alt="gel" width="500">
+<p align="center"><b>Figure 2-2</b>  Colony PCR gel electrophoresis of plasmid pET29(a)-p<i>J23119-SoxR</i>-T-p<i>SoxS</i>-RBS-PhiX174E-T7(1320bp)</p>
+</div>
+<div class="center-img">
+<img src="https://static.igem.wiki/teams/5101/partpage/phix174e.png" alt="cexu" width="600">
+<p align="center"><b>Figure 2-3</b>  plasmid pET29(a)-p<i>J23119-SoxR</i>-T-p<i>SoxS</i>-RBS-PhiX174E-T7 sequencing result</p>
+</div>
+</html>
+==Protein Expression Validation==
+<html>
+We verified the performance of the lysis module through dry experiments and plan to complete wet experimental validation of its lytic function in the future. Through literature review and numerical modeling validation, we found that after inducing the expression of antimicrobial peptides and lysis proteins with NO for twenty minutes, the engineered strain will be lysed by the lysis protein and release the antimicrobial peptides. Mathematical modeling confirmed that at this time, the concentration of antimicrobial peptides is sufficient to reach an effective inhibitory concentration.
+</html>
+==Sequence and Features==
+<partinfo>BBa_K5101004 SequenceAndFeatures</partinfo>
 <!-- Uncomment this to enable Functional Parameter display
-===Functional Parameters===
+==Functional Parameters==
-<partinfo>BBa_K5322002 parameters</partinfo>
+<partinfo>BBa_K5101004 parameters</partinfo>
 <!-- -->