Part:BBa_K2933147

His+Linker f+PEDO-1

This part encodes the fusion protein of His tag and PEDO-1 to promote the expression and purification of target protein(PEDO-1).

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 623
  Illegal PstI site found at 909
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 51
  Illegal PstI site found at 623
  Illegal PstI site found at 909
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 623
  Illegal PstI site found at 909
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 623
  Illegal PstI site found at 909
  
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Biology

This composite part is made up with three basic parts, the HiS tag, the thrombin restriction site and our target protein PEDO-1. It encodes a protein which is PEDO-1 fused with His tag. The fusion protein is about 32.2 kD. It is convenient for us to purify our target protein.

Molecular cloning

First, we used the vector pET28a to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.

Figure 1. Left: The PCR result of PEDO-1. Right: The verification results by enzyme digestion.

References

[1]Dereje Dadi Gudeta, Valeria Bortolaia,The Soil Microbiota Harbors a Diversity of Carbapenem-Hydrolyzing β-Lactamases of Potential Clinical Relevance[J]，Antimicrobial Agents and chemotherapy,January 2016