Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Yin Wu
Date created: 2016-09-13 11:00:00
Date modified: 2016-10-07 02:39:55
SUP35+sfGFP11
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_K2009701_sequence (Version 1) |
Description
Sup35??????PSB1C3 length: 783bpDerived from: PCR from the plasmid provided from Dong Men
introduction
We construted this part as a component of our pro priontein system.
This composite part is contructed by ligating SUP35NM gene and the gene of sfGFP11(with linker).
The split GFP system is simple and does not change fusion proteinsolubility. 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.
Sup35p is a subunit of the translation termination complex. In nature, it has been proved to have kinds of prionic effect on the stress response, When the conformation is transformed into prionic form, the termination of translation is less effective and thus new longer proteins form (True and Lindquist, 2000; Tyedmers et al., 2008). Research found that if the sequence corresponding to the NM domains of Sup35p is fused to other gene, the protein resulting of this construction acquires the prionic behaviour (Li and Lindquist, 2000).
Tyedmers et al. (2000) found that heat shock is a significantly relevant factor to trigger the prionic conformation. Heat shock will induce The cells the prionic phenotype, it is an autocatalytic process and finally, all the protein is found in the prionic conformation. The cells resulting show the phenotype [PSI+]. One of the most possible reason for phenomenon is that HSP104 play an important role in the formation and maintenance of the amyloid fiber (Halfmann et al., 2009).
The SUP35NM gene we used is provided by Dong Men, PhD of Wuhan Insititue of Virology of Chinese Academy of Sciences, so that the sequence is not quite the same as the existing sequence in the Parts Registry, for a lot of mutations have been done. The standardization of this gene is did by ourselves by adding the Biobrick prefix and suffix to its ends and doing a site mutation to eliminate the PstI cutting site inside it.
sequence
ATGGGCGATTCAAACCAAGGCAACAATCAGCAAAACTACCAGCAATACAGCCAGAACGGTAACCAACAACAAGGTAACAACAGATACCAAGGTTATCAAGCTTACAATGCTCAAGCCCAACCTGCcGGTGGGTACTACCAAAATTACCAAGGTTATTCTGGGTACCAACAAGGTGGCTATCAACAGTACAATCCTCAAGGCGGTTATCAGCAGCAATTCAATCCACAAGGTGGCCGTGGAAATTACAAAAACTTCAACTACAATAACAGTTTGCAAGGATATCAAGCTGGTTTCCAACCACAGTCTCAAGGTATGTCTTTGAACGACTTTCAAAAGCAACAAAAGCAGGCCGCTCCCAAACCAAAGAAGACTTTGAAGCTTGTCTCCAGTTCCGGTATCAAGTTGGCCAATGCTACCAAGAAGGTTGACACAAAACCTGCCGAATCTGATAAGAAAGAGGAAGAGAAGTCTGCTGAAACCAAAGAACCAACTAAAGAGCCAACAAAGGTCGAAGAACCAGTTAAAAAGGAGGAGAAACCAGTCCAGACTGAAGAAAAGAAGGAGGAAAAATCGGAACTTCCAAAGGTAGAAGACCTTAAAATCTCTGAATCAACAGATAATACCAACAATGCCAATGTTACCAGTGCTGATGCCTTGATCAAGGAACAGGAAGAAGAAGTGGATGACGAAGTTGTTAACGATtaCTAGATGgatggagggtctggtggcggatcaagagaccacatggtccttcatgagtatgtaaatgctgctgggattaca
(All the sequence has been testified by Sangon)