Difference between revisions of "Part:BBa K4759203"

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<partinfo>BBa_K4759203 short</partinfo>
 
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we selected high-copy plasmids pRSFDuet, medium-high copy plasmids pETDuet and pET28a, and low-copy plasmid pACYCDuet as carriers to express Olep in E. coli, and the results showed that the recombinant strains constructed with high-copy plasmid pRSFDuet as the carrier had the highest expression of p450 enzyme, cell growth rate, and substrate conversion rate.
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we selected high-copy plasmids pRSFDuet, medium-high copy plasmids pETDuet and pET28a, and low-copy plasmid pACYCDuet as carriers to express Olep in E. coli, and the results showed that the recombinant strains constructed with high-copy plasmid pRSFDuet as the carrier had the highest expression of p450 enzyme, cell growth rate, and substrate conversion rate. T7-RBS4-Olep is expressed by the plasmid vector pETDuet/pRSFDuet.
  
  

Latest revision as of 07:11, 10 October 2023


T7-RBS4-OleP

we selected high-copy plasmids pRSFDuet, medium-high copy plasmids pETDuet and pET28a, and low-copy plasmid pACYCDuet as carriers to express Olep in E. coli, and the results showed that the recombinant strains constructed with high-copy plasmid pRSFDuet as the carrier had the highest expression of p450 enzyme, cell growth rate, and substrate conversion rate. T7-RBS4-Olep is expressed by the plasmid vector pETDuet/pRSFDuet.


Usage and Biology

OleP, the cytochrome P450 epoxidase from Streptomyces antibioticus involved in Oleandomycin biosynthesis: functional analysis and crystallographic structure in complex with clotrimazole. PDB DOI: https://doi.org/10.2210/pdb4XE3/pdb Classification: OXIDOREDUCTASE Organism(s): Streptomyces antibioticus Expression System: Escherichia coli BL21(DE3)

4xe3.png

we selected high-copy plasmids pRSFDuet, medium-high copy plasmids pETDuet and pET28a, and low-copy plasmid pACYCDuet as carriers to express Olep in E. coli, and the results showed that the recombinant strains constructed with high-copy plasmid pRSFDuet as the carrier had the highest expression of p450 enzyme, cell growth rate, and substrate conversion rate.

2-1.png

Fig1: Selection of suitable plasmid. WT represents E. coli O1 strain. The blue-filled triangle represents the biomass (OD600). The red hollow triangle represents the conversion rate (%). Values and triangles represent the means and standard deviations of biological triplicates.

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 102
    Illegal AgeI site found at 200
  • 1000
    COMPATIBLE WITH RFC[1000]