Part:BBa_K3784001
fldB (Encoding 2-hydroxyacyl-CoA dehydratase protein family)
The fldB is from the FLD gene cluster of Clostridium sporogenes, involved in the conversion of tryptophan to 3-Indolepropionic acid. In the five-step metabolic pathway we reconstructed, fldB encodes subunit of 2-hydroxyacyl-CoA dehydratase family protein, combined with another subunit encoded by fldC to become 2-hydroxyacyl-CoA dehydratase family protein, which catalyze indole lactic acid to produce indole acrylic acid.
Characterization
We successfully inserted the fldB gene into the His-6p-MBP-RSFD plasmid and transformed it into E. coli BL21(DE3) and induced it to express. After extracting total RNA, we obtained its cDNA by reverse transcription. We used cDNA as a template to amplify the fragment, then checked it by agarose gel electrophoresis. We chose rsmA (16S rRNA m(6)2A1518, m(6)2A1519 dimethyltransferase) as the internal reference gene and set a negative control. The fldB is well expressed and the expression levels of internal reference genes in the experimental group (rsmAE) and the control group (rsmAC) were consistent.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1169
Illegal EcoRI site found at 1241 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1169
Illegal EcoRI site found at 1241 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1169
Illegal EcoRI site found at 1241
Illegal BglII site found at 48 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1169
Illegal EcoRI site found at 1241 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1169
Illegal EcoRI site found at 1241 - 1000COMPATIBLE WITH RFC[1000]
| None |