Part:BBa_K2197302
Resulting repressor is chimeric mammalian urate-dependent transsilencer (mUTS).
Design of BBa_K2197300
Expression of a strong repressor (mUTS)
Operator site and GFP
Expression without uric acid (-UA)
mUTS binds to HucO and repress the expression of downstream GFP.
expression with uric acid (+UA)
mUTS dissociates with HucO to different extents according to the concentration of uric acid, thus expressing downstream
GFP at different levels.
BBa_K2197302 - expressing repressor protein with KRAB amplification which the resulting repressor is chimeric mammalian urate-dependent transsilencer (mUTS). This is a subpart of BBa_K2197300. Engineered E.coli encodes part BBa_K2197302, which expresses a repressor protein with KRAB amplification. Bacterial transcriptional repressor (HucR) was engineered to be a stronger repressor by fusing it to the C terminus of the Krueppel-associated box (KRAB) protein domain15. The resulting repressor is a chimeric mammalian urate-dependent transsilencer (mUTS). HucR binds to a DNA sequence motif (hucO) in the absence of uric acid. When uric aicd is present, HucR dissociates from DNA, thereby allows the expression of a downstream gene. According the research, the expression of the downstream gene is regulated by the concentration of uric acid.
When this part is put in BBa_K2197300, it works with BBa_K2197303 thus amplifies the repression according to the concentration of uric acid, that is, the higher the concentration, the weaker the repression is.
When this part is put in a composite part with BBa_K2197303, the gene's expression will be regulated by the concentration of uric acid.
Bellefroid, E.J., Poncelet, D.A., Lecocq, P.J., Revelant, O. & Martial, J.A. The evolutionarily conserved Kruppel-associated box domain defines a subfamily of eukaryotic multifingered proteins. Proc. Natl. Acad. Sci. USA 88, 3608–3612 (1991).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 508
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 244
Illegal SapI.rc site found at 208
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