Difference between revisions of "Part:BBa K5236025"
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<partinfo>BBa_K5236025 short</partinfo>
 
<partinfo>BBa_K5236025 short</partinfo>
  
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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.
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The sequence of BhrPETase was identified by the Shingo group in a metagenomic study on uncultured thermophiles, and was deposited into the NCBI database by the group in 2018 and annotated as a PET hydrolase [1]. This basic part encoding the BhrPETase, which has been predicted and optimized by Wu et al. And was constructed and modified as WT BhrPETase in our project.
  
 
===Usage and Biology===
 
===Usage and Biology===
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<center>Fig.1 The DNA gel electrophoresis result </center>
 
<center>Fig.1 The DNA gel electrophoresis result </center>
  
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<center><html><img src ="" width = "50%"><br></html></center>
 
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<center>Fig.2 The result of DNA sequencing  </center>
 
  
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When we had completed the plasmid construction and transformation. We need to construct and test the enzymatic activity.   
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When we had completed the plasmid construction and transformation. We need to construct and test the BhrPETase activity.   
 
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Revision as of 13:35, 1 October 2024


BhrPETase

The sequence of BhrPETase was identified by the Shingo group in a metagenomic study on uncultured thermophiles, and was deposited into the NCBI database by the group in 2018 and annotated as a PET hydrolase [1]. This basic part encoding the BhrPETase, which has been 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


When we had completed the plasmid construction and transformation. We need to construct and test the BhrPETase 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]