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 subunit of 2-hydroxyacyl-CoA dehydratase family protein, combined with another subunit encoded by fldB to become 2-hydroxyacyl-CoA dehydratase family protein, which catalyze indole lactic acid to produce indole acrylic acid.
Characterization
We successfully inserted the fldC 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 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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