Difference between revisions of "Part:BBa K5392021"

Latest revision as of 14:56, 30 September 2024

Vector-ZaTdT-K337A-KanR

Description

To obtain the desired saturation mutagenesis, oligonucleotide primers were designed with degenerate codons. In addition, each single-site saturation mutant was generated according to the PCR-based QuickChange method. The PCR was performed according to the operation manual. The PCR product was digested with DpnI restriction enzyme and transformed into E.coli DH5-alpha competent cells.We will extract the plasmid and sequence it to make sure the mutation was successful.

Experiment

SDS-PAGE of ZaTdT-K337A

We transfected the Sequencing is correct ZaTdT-K337A plasmid into E.coli BL21(DE3) competent cell. After overnight, an appropriate colony was used to express the mutant protein and verify its activity

Catalytic activity assay of ZaTdT-K337A

We transfected the Sequencing is correct ZaTdT-K337A plasmid into E.coli BL21(DE3) competent cell. After overnight, an appropriate colony was used to express the mutant protein and verify its activity

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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+===<strong> Catalytic activity assay of ZaTdT-K337A</strong>===
+We transfected the Sequencing is correct ZaTdT-K337A 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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