Difference between revisions of "Part:BBa K5392015"
 
(One intermediate revision by the same user not shown)
Line 11: Line 11:
  
 
</style><div id="a1">
 
</style><div id="a1">
−
<img src="https://static.igem.wiki/teams/5392/pet28a-zatdt-page.png" width="407" height="262"/>
+
<img src="https://static.igem.wiki/teams/5392/pet28a-zatdt-page-1.png" width="407" height="262"/>
 
</div></html>
 
</div></html>
  
Line 25: Line 25:
 
</style><div id="a2">
 
</style><div id="a2">
 
<img src="https://static.igem.wiki/teams/5392/zatdt.png" width="366" height="201"/>
 
<img src="https://static.igem.wiki/teams/5392/zatdt.png" width="366" height="201"/>
 +
</div></html>
 +
 +
===<strong> Catalytic activity assay of ZaTdT</strong>===
 +
We transfected the Sequencing is correct ZaTdT plasmid into E.coli BL21(DE3) competent cell. After overnight, an appropriate colony was used to express the mutant protein and verify its activity
 +
 +
<html><style>
 +
img{margin:auto;}
 +
#a3{width:388px;height:130px;margin:auto;border:3px solid grey}
 +
 +
</style><div id="a3">
 +
<img src="https://static.igem.wiki/teams/5392/zatdt-denaturing-page.png" width="388" height=130""/>
 
</div></html>
 
</div></html>

Latest revision as of 15:06, 30 September 2024


Vector-ZaTdT-KanR

Description

We incorporated the sequences of the ZaTdT gene into the pET28a vector. Then the vector plasmid was transfected into E.coli DH5-alpha competent cells for purification and amplification. In this way, plasmids with the ZaTdT gene were obtained.


Experiment

SDS-PAGE of ZaTdT

We transfected the ZaTdT gene-bonded pET28a plasmid into E.coli BL21(DE3) competent cell. After overnight, Colonies were then picked and performs protein expression. We identified the ZaTdT expression by SDS-PAGE and tested the incorporation of modified nucleotides by purified ZaTdT with polyacrylamide gel electrophoresis.

Catalytic activity assay of ZaTdT

We transfected the Sequencing is correct ZaTdT plasmid into E.coli BL21(DE3) competent cell. After overnight, an appropriate colony was used to express the mutant protein and verify its activity