Part:BBa_K2282008

BBa_2282001 (AmilCP+Ds box) supplemented with BBa_K2282002 (CspA 5'UTR)

Usage and Biology

The DSBox (Downstream Box) sequence increases the translation of the mRNA at low temperature. At low temperatures translation of most transcripts is strongly reduced. 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). The 5’ UTR contains a regulatory sequence called “Cold Box”​ that plays an important role in the stabilisation of the mRNA at low temperature (ATTAAA) (Mitta et al, 1997). The structure of this 5’UTR is extremely unstable at high temperature, as it is believed to be hydrolysed by an RNAse, leading to an average lifetime of 12 seconds stability at 37°C, as opposed to 20 minutes at 15°C, a temperature at which the RNAse is no longer active (Barria et al. 2013). These two elements are parts of the cold shock response machinery. This sequence was used to check the effect of 5’UTR and DSBox on the expression of AmilCP, a coral blue chromoprotein under a strong Anderson constitutive promoter (BBa_J23100).

Source of this part

Please refer to the parts BBa_K2282006 and BBa_K2282007. Sequences of the 5’UTR, DSBox and other cold-inducing 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 design consideration as the parts BBa_K2282006 and BBa_K2282007. Sequences of the 5’UTR, DSBox and other cold-inducing elements are very close to the parts of the iGEM10_Mexico-UNAM-CINVESTAV that can be found here : http://partsregistry.org/Part:BBa_K328003

References

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

C. Barria et al, “Bacterial adaptation to cold”, Microbiology (2013), 159, 2437–2443

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