Part:BBa_K2924000

Pcpc560 Promoter

A strong promoter for protein expression in cyanobacteria. It is made up of two predicted promoters and 14 transcription factor binding sites, as well as an RBS. The sequence stems from the cpcB gene from Synechocystis sp. PCC 6803

Usage and Biology

The Promoter Pcpc560 was discovered in 2013 by Jie Zhou et al.¹ at the Chinese academy of sciences of Beijing. The name derives from the cyanobacterial gene that it originally expresses, phycocyanin B-subunit (cpcB). Zhou et al used a region of 560 bases upstream of this gene, which contains two predicted promoters and 14 predicted transcription factor binding sites (TFBSs). Negative transcription TFBSs are usually located between 500-1000bp upstream of the gene, so this region was left out of the Pcpc560 construct¹. The promoter carries an RBS, so it can be cloned directly in front of the gene of interest. They tested the promoter for the expression of two genes with enzymatic activity, the native enzyme crotonyl-CoA-specific trans-enoyl-CoA reductase (Ter) and D-lactate dehydrogenase from E. coli K-12. In both cases, protein yields of 15% of soluble protein were reported. If the promoter found by Zhou et al. makes these high yields possible for any expressed protein, cyanobacteria might become a viable chassis for protein production, especially since they do not need any carbon source and do not produce CO2 emissions. The promoter is already present in the registry as BBa_K1968001 and was submitted by the team of Edinburgh in 2016, which was not used in Synechocystis. However, BBa_K1968001 differs slightly from this promoter in its sequence and since the function of a promoter is strictly sequence-dependant, this Pcpc560 promoter is submitted as a new part.

References

https://parts.igem.org/Part:BBa_K1968001 1: Zhou, Jie, et al. "Discovery of a super-strong promoter enables efficient production of heterologous proteins in cyanobacteria." Scientific reports 4 (2014): 4500.

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]