Part:BBa_K1758370

BlcR, repressor of blcABC operon, with promoter and RBS

Usage and biology

BlcR, a protein from Agrobacterium tumefaciens, represses the blcABC-operon. The latter enables the bacterium to grow on γ-butyrolactone as carbon and energy source as depicted in the following figure (Pan et al. 2013).

The Blc-Operon and the catabolism of GBL in A. tumefaciens. GHB: γ-hydroxybutyrate; GBL: γ-butyrolactone; SSA: Succinic semialdehyde; SA: Succinate.

Characterization – BlcR function

We performed EMSA-shifts and verified: BlcR binds to the operator site described in Pan et al. 2013.

EMSA shifts caused by addition of BlcR protein (see BBa_K1758370) and BlcR-sfGFP fusion protein (see BBa_K1758204), respectively, to Cy3-labeled blc-operator site. 5 pmol of following proteins were applied: 1: BlcR-sfGFP, 2: ArsR-sfGFP as negative control (see BBa_K1758203), 3: BlcR, 4: none.

A BlcR-sfGFP fusion protein is also able to bind the blc-operator in BBa_K1758376, as depicted in the following figure.

EMSA shift caused by addition of different amounts of the indicated protein to 0.05 pmol Cy3-labeled operator site. +/- refers to the presence of dithiothreitol in the reaction.

References

Pan, Yi; Wang, Yi; Fuqua, Clay; Chen, Lingling (2013): In vivo analysis of DNA binding and ligand interaction of BlcR, an IclR-type repressor from Agrobacterium tumefaciens. In: Microbiology (Reading, England) 159 (Pt 4), S. 814–822. DOI: 10.1099/mic.0.065680-0.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  Illegal NheI site found at 812
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 520
  Illegal NgoMIV site found at 761
  
• 1000
  COMPATIBLE WITH RFC[1000]