Part:BBa_K2924043

Hpall promoter

The P_HpaII promoter¹, is a strong constitutive promoter that is commonly used for expression in gram-positive bacteria. Since the promoter is not mapped, the sequence includes the 5'UTR, as well as the RBS.

Usage and Biology

The promoter has better affinity to the binding site of RNA polymerase or other transcription activators which help in increasing the transcription rate of downstream DNA to messenger RNA^2,3,4. The P_HpaII promoter stimulates counterclockwise RNA synthesis⁵. Comparable expression strength to that of P₄₃². In our case it was used to produce high amounts of heterologous protein (BBa_K2924033, BBa_K2924052) that is secreted into the culture supernatant.

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]

References

[1]: Westers, L., Dijkstra, D.S., Westers, H., van Diji, J.M., Quax, W.J. (2006). "Secretion of functional human interleukin-3 from Bacillus subtilis." Journal of biotechnology 123.2 (2006): 211-224.

[2]: Liu S. L., Du K. (2012). Enhanced expression of an endoglucanase in Bacillus subtilis by using the sucrose-inducible sacB promoter and improved properties of the recombinant enzyme. Protein Expr Purif. 2012 Jun;83(2):164-8. doi: 10.1016/j.pep.2012.03.015. Epub 2012 Apr 4

[3]: Guan C., Cui W., Cheng J., Zhou L., Guo J., Hu X., Xiao G., Zhou Z. (2015). Construction and development of an auto-regulatory gene expression system in Bacillus subtilis. Microb Cell Fact. 2015; 14: 150. doi: 10.1186/s12934-015-0341-2.

[4]: Chen, Jingqi, et al. "Enhanced extracellular production of α-amylase in Bacillus subtilis by optimization of regulatory elements and over-expression of PrsA lipoprotein." Biotechnology letters 37.4 (2015): 899-906.

[5]: Zhang, Kang, et al. "High-level extracellular protein production in Bacillus subtilis using an optimized dual-promoter expression system." Microbial cell factories 16.1 (2017): 32.