Part:BBa_K1137002
M. Smegmatis FdxA
This biobrick encodes a 7Fe ferredoxin which is an orthologue of ferredoxin FdxC in M. tuberculosis. It contains both one 3Fe-4S and one 4Fe-4S cluster. FdxA is the preferred substrate of FprA and is required for the proper functioning of SirA for sulfite reduction during cysteine metabolism. Thus it required for growth on minimal media. We were able to obtain growth from an E. coli strain lacking native cysI in M9 media supplemented with glucose as a carbon source when co-expressed with BBa_K1137000 and BBa_K1137001. Codon optimized for expression in E. coli.
Growth curves: BL21 (DE3) ΔcysI containing the MycoSIR pathway (MycoSIR E.coli) were grown in liquid minimal media containing Various concentration of IPTG. (A) Replicates of each strain were measured for absorbance in a spectrophotometer every 10 minutes for 14 hours. Growth was observed for the WT BL21 E.coli, (blue), and the MycoSIR E.coli (red). No growth was detected for uninduced MycoSIR E.coli (purple) or for the BL21 (DE3) ΔcysI that did not contain the synthetic pathway (Orange) . (B) Mean Final ODs of all replicates, measured after 14 hours of growth. Growth was detected in zmSIR E.coli and WT BL21 but not in uninduced zmSIR strain.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 144
- 1000COMPATIBLE WITH RFC[1000]
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