dszBBAC with tac promoter and dszD with lac promotor induced by IPTG
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) The T7 RNA polymerase controled system is PCRed from E.coli Bl21.
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)
dszB (a desulfinase gene), E.coli-optimized
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)
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)
We use PCR to get this sequence from E.coli Bl21.
T7 RNA polymerase will not be expressed without IPTG. When IPTG is added to 1 mM, T7 RNA polymerase will be expressed and if there is protein gene with T7 promoter, it will be greatly expressed by the host cell.
dszBAC with tac promoter and dszD with lac promoter
Each single enzyme gene was synthesized by Generay with E.coli optimized and we constructed them together and suquencing comformed. the sequences can be found in NCBI (GenBank Accession number L37363.1)
We constructed dszBCA genes with tac promoter and dszD gene with lac promoter choose E.coli BL21 as our host cell. When IPTG is added to our system, DszABCD 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 enzyme gene was synthesized by Generay with E.coli optimized and we constructed them together and suquencing comformed. the sequences can be found in NCBI (GenBank Accession number L37363.1)
Double terminator consisting of BBa_B0010 and BBa_B0012
Derived from the TE terminator of T7 bacteriophage between Genes 1.3 and 1.4 <genbank>V01146</genbank>.
Transcription terminator for the E.coli RNA polymerase.
Suggested by Sri Kosuri and Drew Endy as a high efficiency terminator. The 5' end cutoff was placed immediately after the TAA stop codon and the 3' end cutoff was placed just prior to the RBS of Gene 1.4 (before AAGGAG). Use anywhere transcription should be stopped when the gene of interest is upstream of this terminator.
Transcriptional terminator consisting of a 64 bp stem-loop.
Conversion of the iGEM parts registry to SBOL2.1