Difference between revisions of "Part:BBa K1972014"

 
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<partinfo>BBa_K1972014 short</partinfo>
 
<partinfo>BBa_K1972014 short</partinfo>
  
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We constructed dszBCAD genes with T7 promoter and IPTG controled system and choose E.coli as our host cell.T7 RNA polymerase will not be expressed without IPTG. When IPTG is added to 1 mM, T7 RNA polymerase will be expressed and dszBCAD genes with T7 promoter will be greatly expressed by the host cell.  The four enzymes will be successful expressed and these four enzymes convert dibenzothiophene (DBT) undergoes through three successive oxidation steps and one a hydrolytic step to 2-hydroxybiphenyl (2HBP) by the 4s pathway.
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We constructed dszB gene with tac promoter and teminator. DszB is a a desulfinase which can convert 2-(2-hydroxybiphenyl) sulfinate (HBPS) to 2-hydroxybiphenyl (2HBP) during the 4S pathway, in which dibenzothiophene (DBT) undergoes three successive oxidation steps and one a hydrolytic step leading to the formation of 2-hydroxybiphenyl (2HBP)
  
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Each single biodesulfurization enzyme gene was synthesized by Generay and we constructed them together and suquencing comformed. the sequences can be found in NCBI (GenBank Accession number L37363.1)  
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This single monooxygenase gene was synthesized whith E.coli codon optimized by Generay and the sequence was found in NCBI (GenBank Accession number L37363.1)
  
 
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Revision as of 13:01, 19 October 2016


dszBBAC with tac promoter and dszD with lac promotor induced by IPTG

We constructed dszB gene with tac promoter and teminator. DszB is a a desulfinase which can convert 2-(2-hydroxybiphenyl) sulfinate (HBPS) to 2-hydroxybiphenyl (2HBP) during the 4S pathway, in which dibenzothiophene (DBT) undergoes three successive oxidation steps and one a hydrolytic step leading to the formation of 2-hydroxybiphenyl (2HBP)

This single monooxygenase gene was synthesized whith E.coli codon optimized by Generay and the sequence was found in NCBI (GenBank Accession number L37363.1)

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 4052
    Illegal XhoI site found at 5558
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 267
    Illegal NgoMIV site found at 1582
    Illegal NgoMIV site found at 3076
    Illegal AgeI site found at 1104
    Illegal AgeI site found at 2419
    Illegal AgeI site found at 3180
    Illegal AgeI site found at 3463
    Illegal AgeI site found at 4281
    Illegal AgeI site found at 5746
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 864
    Illegal BsaI.rc site found at 2179