Part:BBa_K537001

Theophylline Riboswitch 1-CheZ

This DNA part will encode for an RNA riboswitch senstive to theophylline. The RBS is not exposed in the absence of theophylline. When theophylline is present, it binds to the riboswitch and causes a conformational change which leads to the exposure of the RBS and consequently translation of the adjoining gene. In this case, the gene which will be expressed is CheZ, which is a protein fundamental to bacterial movement.

This theophylline riboswitch (type1) CheZ fusion BioBrick activates the expression of the CheZ gene in a theophylline-dependent fashion. It is composed of a theophylline-sensitive riboswitch clone 8.1 (Topp and Gallivan JACS 2007; <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K249026">BBa_K249026 </a> and K343007 <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K411001">BBa_K411001</a> ) that is detached from its associated translation unit (coding region) and fused to a CheZ gene which has a N-fusion prefix and lacks a stop codon (although a TAG stop codon is provided by the RFC 25 suffix). The riboswitch regulated N-terminal part can be fused to a reporter or other CDS.

CheZ is the chief regulator of the molecular events that lead the counter clockwise rotation of the flagella motor during the chemotaxis signal transduction pathway of E.coli. This counter clockwise flagella motor rotation results in bacterial swimming (instead of tumbling) in the presence of a chemoattractant (in this case, theophylline).

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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]