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==SDS-PAGE analysis of 6×His-CsnBD78Y==
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<h2>SDS-PAGE analysis of 6×His-CsnBD78Y</h2>
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  Purification of the CsnB-D78Y enzyme was achieved using Ni-NTA affinity chromatography, and both the unpurified and purified proteins were confirmed via SDS-PAGE. As shown in the figure below, a distinct band at approximately 30 kDa was observed in the lane corresponding to the unpurified enzyme sample, matching the expected theoretical molecular weight. Following purification, the mutant enzyme showed a profile similar to that of CsnB, with a single band at the same molecular weight as the unpurified enzyme solution.
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  <p>Purification of the CsnB-D78Y enzyme was achieved using Ni-NTA affinity chromatography, and both the unpurified and purified proteins were confirmed via SDS-PAGE. As shown in the figure below, a distinct band at approximately 30 kDa was observed in the lane corresponding to the unpurified enzyme sample, matching the expected theoretical molecular weight. Following purification, the mutant enzyme showed a profile similar to that of CsnB, with a single band at the same molecular weight as the unpurified enzyme solution.</p>
  
 
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Latest revision as of 04:39, 30 September 2024


6His-CsnBD78Y

This part is the rational design of the mutated gene CsnB derived from Marine Bacterium Bacilius SP. BY01 with 6×His tag.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 709
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 718
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 58
    Illegal AgeI site found at 541
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

Plasmid construction

We placed 6×His tag into plasmid PET-28a for protein isolation and purification and tranformed PET-28a-6×His-CsnBD78Y into E. coli BL21 (DE3).

SDS-PAGE analysis of 6×His-CsnBD78Y

Purification of the CsnB-D78Y enzyme was achieved using Ni-NTA affinity chromatography, and both the unpurified and purified proteins were confirmed via SDS-PAGE. As shown in the figure below, a distinct band at approximately 30 kDa was observed in the lane corresponding to the unpurified enzyme sample, matching the expected theoretical molecular weight. Following purification, the mutant enzyme showed a profile similar to that of CsnB, with a single band at the same molecular weight as the unpurified enzyme solution.