Difference between revisions of "Part:BBa K1972009"

Revision as of 13:57, 14 October 2016

dszBCAD with Lac promoter

We constructed dszBCAD genes with lac promoter(a weaker promoter to aviod the generation of inclusion body) and 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 and we constructed them together and suquencing comformed. the sequences can be found in NCBI (GenBank Accession number L37363.1)

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1266
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 488
Illegal BamHI site found at 2070
Illegal XhoI site found at 480
Illegal XhoI site found at 3225
Illegal XhoI site found at 4066
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 195
Illegal NgoMIV site found at 2514
Illegal NgoMIV site found at 2729
Illegal NgoMIV site found at 3033
Illegal NgoMIV site found at 3153
Illegal AgeI site found at 2688
Illegal AgeI site found at 4254
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 2220
Illegal SapI.rc site found at 3537

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K1972009 short</partinfo>
-We constructed dszBCAD genes with T7 promoter and 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.
+We constructed dszBCAD genes with lac promoter(a weaker promoter to aviod the generation of inclusion body) and 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 and we constructed them together and suquencing comformed. the sequences can be found in NCBI (GenBank Accession number L37363.1)