Part:BBa_K3971006

Bidirectional promoter from E. coli EC3132

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Biology

This is a native bidirectional promoter of the csc operon from E. coli EC3132, a strain that can naturally consume sucrose [1].

The csc operon consists of the genes cscA, cscB and cscK. cscA codes for sucrose-6-phosphate hydrolase, which hydrolyses sucrose in glucose and fructose. cscB codes for sucrose permease, a sucrose-proton symporter on bacterial cell membranes. cscK codes for fructokinase, which catalyses the conversion of fructose to fructose-6-phosphate [2].

cscB facilitates the import of sucrose into the bacterium, after which it is hydrolysed into glucose and fructose using the gene product of cscA, and then phosphorylated by the gene product of cscK[1].

The broader csc operon consists of two constituent operons, a cscA operon and a cscBK operon, both of which are under the bidirectional promoter. The cscABK genes are regulated by a repressor coded by cscR, which is under the control of a different promoter[2].

This bidirectional promoter shares a 99.55% sequence alignment with its homologue from E. coli W (verified by NCBI BLAST search), a strain that can also naturally consume sucrose.

csc operon and sucrose consumption pathway in E. coli EC3132. Figure adapted from Bruschi et al. (2012) [1]

Sequence of bidirectional promoter region in the csc operon in E. coli EC3132. "The putative −35 and −10 regions are underlined, as well as the putative ribosome-binding sites (RBS) and the start codons of cscK and cscA. Transcriptional start points (TK) are indicated by letters in boldface italic type, the putative csc operators are boxed, and the putative cAMP-CrpA binding sites are marked with diagonal lines." Figure and image description adapted from Jahreis et al. (2002) [1]

References

[1] Jahreis, K., Bentler, L., Bockmann, J., Hans, S., Meyer, A., Siepelmeyer, J., & Lengeler, J. W. (2002). Adaptation of sucrose metabolism in the Escherichia coli wild-type strain EC3132. Journal of bacteriology, 184(19), 5307–5316. https://doi.org/10.1128/JB.184.19.5307-5316.2002

[2] Bruschi, M., Boyes, S. J., Sugiarto, H., Nielsen, L. K., & Vickers, C. E. (2012). A transferable sucrose utilization approach for non-sucrose-utilizing Escherichia coli strains. Biotechnology advances, 30(5), 1001-1010.