Difference between revisions of "Part:BBa K5291033"

 
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===Usage and Biology===
 
===Usage and Biology===
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AlkB2-Rd45-AdhA is designed to accelerate the degradation rate by improving the absorbing and hydroxylation of PE monomer. In order to express above the enzymes in our chassis， pAB1-AlkB2-Rd45-VHB was constructed.<br><br>
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<html><img width = "700" src="" /></html>(lack)
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<br><b>Fig.1 The map of the plasmid pAB1-AlkB2-Rd45-AdhA.</b><br><br>
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We transferred the constructed plasmid into <i>Escherichia coli</i> DH5αstrain and conducted colony PCR. After obtained the correct result we amplified and extracted the constructed plasmids. Then we transferred these plasmids into <i>Pseudomonas aeruginosa</i> PAO1 strain and <i>E. coli</i> BL21 strain and obtained correct colony PCR results, indicating that we successfully constructed strains containing the plasmid pAB1-AlkB2-Rd45-AdhA.<br><br>
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<html><img width = "350" src="https://static.igem.wiki/teams/5291/images/part-wyn/alkbcyp-colony-pcr.png" /></html>
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<br><b>Fig.2 The <i>P.aeryginosa</i> PAO1 colony PCR results of AlkB2-Rd45-AdhA.</b><br><br>
  
 
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Revision as of 03:10, 2 October 2024


pAB1-alkB2-Rd45-adhA

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Usage and Biology

AlkB2-Rd45-AdhA is designed to accelerate the degradation rate by improving the absorbing and hydroxylation of PE monomer. In order to express above the enzymes in our chassis， pAB1-AlkB2-Rd45-VHB was constructed.

(lack)
Fig.1 The map of the plasmid pAB1-AlkB2-Rd45-AdhA.

We transferred the constructed plasmid into Escherichia coli DH5αstrain and conducted colony PCR. After obtained the correct result we amplified and extracted the constructed plasmids. Then we transferred these plasmids into Pseudomonas aeruginosa PAO1 strain and E. coli BL21 strain and obtained correct colony PCR results, indicating that we successfully constructed strains containing the plasmid pAB1-AlkB2-Rd45-AdhA.


Fig.2 The P.aeryginosa PAO1 colony PCR results of AlkB2-Rd45-AdhA.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 902
    Illegal NgoMIV site found at 2433
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1476