Part:BBa_K1884001

Gal4BD-CIB1

Cryptochrome-interacting basic-helix-loop-helix 1(CIB1) is a protein-coded gene. The product of this gene expression is a basic helix-loop-helix (bHLH) protein which would interact with cryptochrome 2 (CRY2), a blue light stimulated photoreceptor, when exposed to blue light. This part is a Gal4 DNA binding domain fused to C terminus of CIB1.

Usage and Biology

This fusion protein is for use in a yeast-two-hybrid system,and a Gal4 DNA binding domian fused to its C terminus. In order to control DNA transcription by blue light, the system is based on a two-hybrid interaction in which a light-mediated protein brings together two halves of a split transcription factor. CRY2 will disconnected with CIB1 in the dark and halt the DNA transcription.

Figure 1.The diagram of light-mediate-controlled yeast-two-hybrid system

BD-CIB1 is 1533bp in length. Fig. 2 shows the DNA sequence of BD-CIB1 is successfully amplified by PCR from psi-Check2 vector. From this electrophoretogram, we can see the brightness of BD-CIB1 PCR product is rather high compared with DNA Marker, which indicates that the PCR product of BD-CIB1 is in a high concerntration.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 218
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 637
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 137