Difference between revisions of "Part:BBa K5392015"
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<partinfo>BBa_K5392015 short</partinfo>
 
<partinfo>BBa_K5392015 short</partinfo>
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Here, we loaded the genes encoding TdT into plasmid vectors. In this way we were able to over-express the ZaTdT in the Escherichia coli BL21.
 
  
 
==Description==
 
==Description==
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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.
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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.
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<html><style>
 
<html><style>
 
img{margin:auto;}
 
img{margin:auto;}
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#a1{width:600px;height:200px;margin:auto;border:3px solid grey}
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#a1{width:407px;height:262px;margin:auto;border:3px solid grey}
  
 
</style><div id="a1">
 
</style><div id="a1">
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<img src="https://static.igem.wiki/teams/5392/wtzatdt.png" width="585" height="196"/>
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<img src="https://static.igem.wiki/teams/5392/pet28a-zatdt-page.png" width="407" height="262"/>
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</div></html>
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==Experiment==
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===<strong>SDS-PAGE of ZaTdT</strong>===
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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.
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<html><style>
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img{margin:auto;}
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#a2{width:370px;height:205px;margin:auto;border:3px solid grey}
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</style><div id="a2">
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<img src="https://static.igem.wiki/teams/5392/zatdt.png" width="366" height="201"/>
 
</div></html>
 
</div></html>

Revision as of 09:47, 28 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.