Part:BBa_K1972014
dszBBAC with tac promoter and dszD with lac promotor induced by IPTG
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.
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)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 4052
Illegal XhoI site found at 5558 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 864
Illegal BsaI.rc site found at 2179
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