Difference between revisions of "Part:BBa K5392015"
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| + | ===<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 | ||
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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
