Difference between revisions of "Part:BBa K5291003"
 
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In degradation module we constructed pAB1-CYPY96F-VHb to accelerate the degradation rate by improving the absorbing and hydroxylation of PE monomer.<br>
 
In degradation module we constructed pAB1-CYPY96F-VHb to accelerate the degradation rate by improving the absorbing and hydroxylation of PE monomer.<br>
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<html><img width = "600" src="https://static.igem.wiki/teams/5291/images/part-wyn/cyp-plasmid.png" /></html><br>
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<html><img width = "600" src="https://static.igem.wiki/teams/5291/images/part-wyn/pab1-cypy96f-vhb-plasmid.png" /></html><br>
 
<b>Fig.1 The map of plasmid pAB1-CYPY96F-VHb.</b><br><br>
 
<b>Fig.1 The map of plasmid pAB1-CYPY96F-VHb.</b><br><br>
  
 
We transferred the constructed plasmid into <i>Escherichia coli</i> DH5α strain, conducted colony PCR and obtained the correct result.
 
We transferred the constructed plasmid into <i>Escherichia coli</i> DH5α strain, conducted colony PCR and obtained the correct result.
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<html><img width = "300" src="https://static.igem.wiki/teams/5291/images/result/degradation/p-aeryginosa-pao1-colony-pcr-results-of-cypy96f-vhb-and-alkb2-rd45-adha1.png" /></html><br>
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<html><img width = "300" src="https://static.igem.wiki/teams/5291/images/part-wyn/gel-vhb-white.png" /></html><br>
 
<b>Fig.2 P.aeryginosa PAO1 colony PCR results of CYPY96F-VHB.</b><br><br>
 
<b>Fig.2 P.aeryginosa PAO1 colony PCR results of CYPY96F-VHB.</b><br><br>
  
 
Moreover,VHb proteins was successfully expressed in the strains. SDS-PAGE was performed and the following results were obtained.<br>
 
Moreover,VHb proteins was successfully expressed in the strains. SDS-PAGE was performed and the following results were obtained.<br>
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<html><img width = "500" src="https://static.igem.wiki/teams/5291/images/part-wyn/sds-page-cyp.png" /></html><br>
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<html><img width = "500" src="https://static.igem.wiki/teams/5291/images/part-wyn/sds-vhb-white.png" /></html><br>
 
<b>Fig.3 The SDS-PAGE result of VHb.</b><br><br>
 
<b>Fig.3 The SDS-PAGE result of VHb.</b><br><br>
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Since we set CYPY96F and VHB as a whole, the overall functions need moving to the part BBa_K5291034.<br>
+
Since we set CYPY96F and VHb as a whole, the overall functions need moving to the part BBa_K5291034.<br>
  
 
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Latest revision as of 05:06, 2 October 2024


vgb: Vitreoscilla hemoglobin

vgb encodes VHb. VHb, the first discovered bacterial hemoglobin, is a soluble heme-binding protein with a faster rate of oxygen dissociation. It can enhance cell growth, product synthesis and stress tolerance. Especially under oxygen-limited conditions, VHb can interact with terminal oxidase to deliver enough oxygen to improve the degradation ability of CYPY96F on PE.

Usage and Biology

In degradation module we constructed pAB1-CYPY96F-VHb to accelerate the degradation rate by improving the absorbing and hydroxylation of PE monomer.

Fig.1 The map of plasmid pAB1-CYPY96F-VHb.

We transferred the constructed plasmid into Escherichia coli DH5α strain, conducted colony PCR and obtained the correct result.
Fig.2 P.aeryginosa PAO1 colony PCR results of CYPY96F-VHB.

Moreover,VHb proteins was successfully expressed in the strains. SDS-PAGE was performed and the following results were obtained.

Fig.3 The SDS-PAGE result of VHb.

Since we set CYPY96F and VHb as a whole, the overall functions need moving to the part BBa_K5291034.

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]