Part:BBa_K2282010

BBa_K2282006 (AmilCP + Ds box) with BBa_K2282001 (CspA promoter + Up element)

Usage and Biology

This sequence is used to evaluate our cold shock response. The UP element is believed to stimulate the transcription of the CspA gene at a cold temperature, although some reports conclude the opposite (Phadtare et al., 2005). This promoter plays a role in transcription, though does not relate to the translation regulation (DSBox, 5’UTR). The CspA promoter is considered a strong promoter (Mitta et al., 1997). The DSBox is a cis-acting mRNA element enabling assembly of the translation pre-initiation complex, likely through interaction with ribosomal protein S1. (Studer et al 2006, Duval et al, 2013, Qu et al 2012)

Source of this part

Refer to the part BBa_K2282005. Only the addition of the DSBox in the AmilCP coding sequence makes the difference between this part and BBa_K2282005. Sequences of the DSBox and other cold-induced elements are very close to the parts of the iGEM10_Mexico-UNAM-CINVESTAV that can be found here : http://partsregistry.org/Part:BBa_K328003

Design consideration

The same difficulties as BBa_K2282005

References

Phadtare S, Severinov K. Extended −10 Motif Is Critical for Activity of the cspA Promoter but Does Not Contribute to Low-Temperature Transcription. Journal of Bacteriology. 2005;187(18):6584-6589. doi:10.1128/JB.187.18.6584-6589.2005.

Masanori Mitta et al, Deletion analysis of cspA of Escherichia coli requirement of the AT-rich UP element for cspA transcription and the downstream box in the coding region for its cold shock induction, Molecular Microbiology (1997) 26(2), 321–335 Studer, Sean M., and Simpson Joseph. “Unfolding of mRNA Secondary Structure by the Bacterial Translation Initiation Complex.” Molecular Cell 22, no. 1 (April 7, 2006): 105–15. doi:10.1016/j.molcel.2006.02.014.

Duval, Mélodie, Alexey Korepanov, Olivier Fuchsbauer, Pierre Fechter, Andrea Haller, Attilio Fabbretti, Laurence Choulier, et al. “Escherichia Coli Ribosomal Protein S1 Unfolds Structured mRNAs Onto the Ribosome for Active Translation Initiation.” PLOS Biology 11, no. 12 (December 10, 2013): e1001731. doi:10.1371/journal.pbio.1001731.

Qu, Xiaohui, Laura Lancaster, Harry F. Noller, Carlos Bustamante, and Ignacio Tinoco. “Ribosomal Protein S1 Unwinds Double-Stranded RNA in Multiple Steps.” Proceedings of the National Academy of Sciences 109, no. 36 (September 4, 2012): 14458–63. doi:10.1073/pnas.1208950109.

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]