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===Usage and Biology=== | ===Usage and Biology=== | ||
Revision as of 10:20, 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.
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.
Usage
To make this parts RFC 10 competible, we use Site-directed mutagenesis to mutate the old sequence so that none of these four Restriction enzyme(EcoR1, Xba1， Sep1， Pst1) can digest this gene.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 218
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 637
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 137