Difference between revisions of "Part:BBa K1758370"

Revision as of 05:03, 19 September 2015

BlcR, repressor of blcABC operon, with promoter and RBS

Usage and biology

BlcR, a gene of 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 BlcR and BlcR-sfGFP — 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.

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]

@@ Line 27: / Line 27: @@
      </figure>
+</br>
+<h3>References</h3>
+<p id="Pan2013">
+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.
+</p>