Part:BBa_K3784002
fldC (Encoding 2-hydroxyacyl-CoA dehydratase family protein)
The fldC 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, fldC encodes encodes subunit of 2-hydroxyacyl-CoA dehydratase protein, combined with another subunit encoded by fldB to become 2-hydroxyacyl-CoA dehydratase protein, which catalyze indole lactic acid to produce indole acrylic acid.
Characterization
We successfully inserted the fldC gene into the plasmid, which is transcribed by a self-contained promoter cloned from Clostridium sporogenes.
We transformed the plasmid 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 fldC 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 XbaI site found at 799
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 799
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 799
- 1000COMPATIBLE WITH RFC[1000]
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