Difference between revisions of "Part:BBa K4122020"

 
 
(3 intermediate revisions by the same user not shown)
Line 2: Line 2:
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K4122020 short</partinfo>
 
<partinfo>BBa_K4122020 short</partinfo>
 +
<html>
  
−
3 PARTS
+
<p>This parts is composed of 3 PARTS:</p>
−
PFBA1-SP-PETase-His-Spytag-T-ADH2
+
<p>PFBA1-SP-PETase-His-Spytag-T-ADH2</p>
−
PFBA1-SP-MHETase-cMyc-Snooptag-T-PGK1
+
<p>PFBA1-SP-MHETase-cMyc-Snooptag-T-PGK1</p>
−
SP-CBM-SC-SC-SNC-SC-V5-7813
+
<p>SP-CBM-SC-SC-SNC-SC-V5-7813</p>
  
−
In this assembly, SP is signal peptide, the same as last year (BBa_K4122014). GFP is replaced by PETase. His-tag is used for protein purification, and Spytag will form a tight connection with spycatcher, which in turn anchors the attached PETase to the cell surface.Promoter-PFBA1, Terminator-TADH2. RFP is replaced by MHETase. His-tag is used for protein purification, and Snooptag will form a tight connection with snoopcatcher, which in turn anchors the attached MHETase to the cell surface.Promoter-PFBA1, Terminator-T-PGK1.
+
<p>When replacing RFP and GFP with MHETase and PETase, we did not observe immunofluorescence with secondary antibodies that should theoretically bind specifically to the V5 tag.</p>
 +
<p>Then we started to analyze.</p >
 +
 
 +
<img src="https://static.igem.wiki/teams/4122/wiki/parts/10-the-construction-of-plasmid-ts-pgapdh-teno1a-the-surface-display-system-for-displaying-both-petase-and-mhetase.png" style = "width:70%;">
 +
<br/><b>Fig.1</b> The construction of plasmid Ts-PGAPDH--TENO1A, the surface display system for displaying both PETase and MHETase.
 +
<br/>
  
  

Latest revision as of 15:05, 8 October 2022


SP-MHETase-cMyc-Snooptag+SP-PETase-His-Spytag+SP-CBM-SC-SC-SNC-SC-V5-7813

This parts is composed of 3 PARTS:

PFBA1-SP-PETase-His-Spytag-T-ADH2

PFBA1-SP-MHETase-cMyc-Snooptag-T-PGK1

SP-CBM-SC-SC-SNC-SC-V5-7813

When replacing RFP and GFP with MHETase and PETase, we did not observe immunofluorescence with secondary antibodies that should theoretically bind specifically to the V5 tag.

Then we started to analyze.


Fig.1 The construction of plasmid Ts-PGAPDH--TENO1A, the surface display system for displaying both PETase and MHETase.
Sequence and Features BBa_K4122020 SequenceAndFeatures