Difference between revisions of "Part:BBa K1884001"

(Usage and Biology)
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<figure style="text-align: center"><img style="width:60%" src="https://static.igem.org/mediawiki/2016/e/e0/Jiaomu2.png"/><figcaption style="text-align:center"><b>Figure 2.</b> Electrophoretic analysis of PCR product of hTERT promoter from pSB1C3. <figcaption style="text-align:center">(1:DL2000 DNA Marker 2:PCR product)</figcaption></figure>
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<figure style="text-align: center"><img style="width:60%" src="https://static.igem.org/mediawiki/2016/e/e0/Jiaomu2.png"/><figcaption style="text-align:center"><b>Figure 1.</b> Electrophoretic analysis of PCR product of hTERT promoter from pSB1C3. <figcaption style="text-align:center">(1:DL2000 DNA Marker 2:PCR product)</figcaption></figure>
  
 
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Revision as of 16:51, 16 October 2016


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. Electrophoretic analysis of PCR product of hTERT promoter from pSB1C3.
(1:DL2000 DNA Marker 2:PCR product)


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