Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Yin Wu
Date created: 2016-09-13 11:00:00
Date modified: 2016-09-15 02:33:25
sfGFP11
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K2009820_sequence (Version 1) |
Description
sfGFP11 length: 48bpDerived from:synthesis from Sangon
sfGFP11??????PSB1C3 is an expression plasmid which insert sfGFP11 into PSB1C3.
before sfGFP11, we add a linker(gatggagggtctggtggcggatca) to achieve our goal.SfGFP11 is a part of GFP(from 214bp to 230bp), GFP has been mutated to improve its solubilityand self-associating activity. When it express, it will emit green fluorescenceslightly under the fluorescence microscope.
We try to find anideal protein tag to be work both invivo and invitro and it can provide a sensitive measurable signalwhich don???t need external chemical reagents or substrates. Finally we find away to accomplish this goal?????? dividing GFP into sfGFP1-10and sfGFP11. Either the sfGFP1-10 or sfGFP11 will emit green fluorescence slightly under the fluorescencemicroscope. However, when sfGFP1-10 and sfGFP11 express insame cell, they will interact each other and emit more intense fluorescence thaneach of them. The split GFP system is simple and does not change fusion proteinsolubility.
Usage and biology:The split GFP system has manypractical applications. Obtaining soluble, well-folded recombinant proteins fordownstream applications requires screening large numbers of protein variants (mutants,fragments, fusion tags, folding partners) and testing many expression orrefolding conditions.(Ste??phanieCabantous, Thomas C Terwilliger & Geoffrey S Waldo,2005)
Part sequence
agagaccacatggtccttcatgagtatgtaaatgctgctgggattaca
(All the sequence has been testified by Sangon)
Notes
We give up PCR from the gene of GFP due to the fact that to achieve the PCR, GFP gene should be mutated too many times. It is more convinent to chenical synthese directly.Source
chemical synthesing sequence which is designed by ???Protein tagging and detection with engineeredself-assembling fragments of green fluorescent protein Ste??phanie Cabantous, Thomas C Terwilliger & Geoffrey S Waldo，2005???