Difference between revisions of "Part:BBa K4759222"

Revision as of 08:56, 12 October 2023

T7-RBS1-Olep

(pRSFDuet/pACYCDuet) expresses redox chaperone, and pRSFDuet expresses GFP1-10. We used pRSFDuet / pACYCDuet as the plasmid vector, T7-RBS1 to express the redox chaperone, or we used pRSFDuet as the plasmid vector ，T7-RBS1 to express GFP1-10.

Usage and Biology

We selected four conventional redox partners (BM3, CamA/CamB, SelFdR0978/SelFdx1499, PetH/PetF) in combination with the P450 enzyme. Four groups of redox partners were constructed on the high-copy plasmid pRSFDuet to obtain recombinant plasmids: pRSFDuet-BM3-olep, pRSFDuet-camA-camB-olep, pRSFDuet-FdR0978-Fdx1499-olep, and pRSFDuet-petH-petF-olep. Then they were transformed to C41 (DE3) to obtain the recombinant strain R2 to R5. The recombinant strains R2 to R5 were subjected to shaker fermentation experiments.
The recombinant strain R5 (containing recombinant plasmid pRSFDuet-petH-petF-olep) had the highest conversion rate, which significantly increased from 41.4% to 85.6%. Therefore, the redox partners PetH/PetF derived from Synechocystis was successfully screened as the most suitable redox partners for the P450 enzyme Olep, and the construction of the sfGFP sensor was verified, which could efficiently and accurately screen the redox partner adapted by the P450 enzyme.

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 AgeI site found at 103
Illegal AgeI site found at 201
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K4759222 short</partinfo>
-(pRSFDuet/pACYCDuet) expresses redox chaperone, and pRSFDuet expresses GFP1-10. We used pRSFDuet / pACYCDuet as the plasmid vector, T7-RBS1 to express the redox chaperone, or we used pRSFDuet as the plasmid vector ，T7-RBS1 to express GFP1-10
+(pRSFDuet/pACYCDuet) expresses redox chaperone, and pRSFDuet expresses GFP1-10. We used pRSFDuet / pACYCDuet as the plasmid vector, T7-RBS1 to express the redox chaperone, or we used pRSFDuet as the plasmid vector ，T7-RBS1 to express GFP1-10.
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+We selected four conventional redox partners (BM3, CamA/CamB, SelFdR0978/SelFdx1499, PetH/PetF) in combination with the P450 enzyme. Four groups of redox partners were constructed on the high-copy plasmid pRSFDuet to obtain recombinant plasmids: pRSFDuet-BM3-olep, pRSFDuet-camA-camB-olep, pRSFDuet-FdR0978-Fdx1499-olep, and pRSFDuet-petH-petF-olep. Then they were transformed to C41 (DE3) to obtain the recombinant strain R2 to R5. The recombinant strains R2 to R5 were subjected to shaker fermentation experiments. <br>
+The recombinant strain R5 (containing recombinant plasmid pRSFDuet-petH-petF-olep) had the highest conversion rate, which significantly increased from 41.4% to 85.6%. Therefore, the redox partners PetH/PetF derived from Synechocystis was successfully screened as the most suitable redox partners for the P450 enzyme Olep, and the construction of the sfGFP sensor was verified, which could efficiently and accurately screen the redox partner adapted by the P450 enzyme.
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