Difference between revisions of "Part:BBa K3040014"

 
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pFadD promoter is one of the regulator in the enzymes of fatty acid biosynthesis in E. coli. It is composed of two fadR recognition sites, and one CRP binding site. The fatty acid metabolism system of E. coli is consist of many parts, fadD, fadL, fadR, fadA, etc. Interestingly, each enzyme in this family has its own different sequence of fadR recognition site in its promoter, which makes these promoter having different strength, yet can still be interchangeable for us to apply in our system.  
 
pFadD promoter is one of the regulator in the enzymes of fatty acid biosynthesis in E. coli. It is composed of two fadR recognition sites, and one CRP binding site. The fatty acid metabolism system of E. coli is consist of many parts, fadD, fadL, fadR, fadA, etc. Interestingly, each enzyme in this family has its own different sequence of fadR recognition site in its promoter, which makes these promoter having different strength, yet can still be interchangeable for us to apply in our system.  
 
Based on this fact, we tried pFadD promoter to take place of the weak pFadBA promoter. With two fadR recognition sites native in pFadD, we assume a better result in the sensitivity and a lower leakage in our pFadD promoter. Yet, we further modified pFadD promoter by replacing its CRP binding site with a FadR recognition site of the sequence of pFadBA promoter.
 
Based on this fact, we tried pFadD promoter to take place of the weak pFadBA promoter. With two fadR recognition sites native in pFadD, we assume a better result in the sensitivity and a lower leakage in our pFadD promoter. Yet, we further modified pFadD promoter by replacing its CRP binding site with a FadR recognition site of the sequence of pFadBA promoter.
 +
==Reference==
 +
FENG, Youjun; CRONAN, John E. Crosstalk of Escherichia coli FadR with global regulators in expression of fatty acid transport genes. PloS one, 2012, 7.9: e46275.
  
  

Revision as of 19:15, 18 October 2019


pFadD promoter with pLac regulating downstream RFP

pFadD promoter is one of the regulator in the enzymes of fatty acid biosynthesis in E. coli. It is composed of two fadR recognition sites, and one CRP binding site. The fatty acid metabolism system of E. coli is consist of many parts, fadD, fadL, fadR, fadA, etc. Interestingly, each enzyme in this family has its own different sequence of fadR recognition site in its promoter, which makes these promoter having different strength, yet can still be interchangeable for us to apply in our system. Based on this fact, we tried pFadD promoter to take place of the weak pFadBA promoter. With two fadR recognition sites native in pFadD, we assume a better result in the sensitivity and a lower leakage in our pFadD promoter. Yet, we further modified pFadD promoter by replacing its CRP binding site with a FadR recognition site of the sequence of pFadBA promoter.

Reference

FENG, Youjun; CRONAN, John E. Crosstalk of Escherichia coli FadR with global regulators in expression of fatty acid transport genes. PloS one, 2012, 7.9: e46275.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 30
    Illegal suffix found in sequence at 230
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 30
    Illegal SpeI site found at 231
    Illegal PstI site found at 245
    Illegal NotI site found at 36
    Illegal NotI site found at 238
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 30
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 30
    Illegal suffix found in sequence at 231
  • 25
    INCOMPATIBLE WITH RFC[25]
    Unknown
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Unknown