Part:BBa_K5236025

BhrPETase

This basic part encoding the BhrPETase, which has been sequence predicted and optimized by Wu et al. And was constructed and modified as WT BhrPETase in our project.

Usage and Biology

To insert the parts into plasmids, we’ve designed primers and performed PCRs. Then, our genes were recombined into plasmids and transformed into chassis. By conducting colony PCR, we are able to test if our parts have been transformed into E.coli successfully. The following result of electrophoresis proves that we’ve inserted genes into chassis since the sequence containing our mutated genes has a total of 798 base pairs and the results are in the right location.

Fig.1 The DNA gel electrophoresis result
Fig.2 The result of DNA sequencing

When we had completed the plasmid construction and transformation. We need to construct and test the enzymatic activity. .

Fig.3 Mutated BhrPETase Dynamic Curve
Fig.4 Protein electrophoresis result

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]