WebbThe intact PhoCl-MBP fusion (75 kDa) was eluted first from the column at a volume of ∼47 mL. This was followed by the cleaved peptide-MBP fusion (45 kDa) at ∼52 mL and then … WebbFig. S5 Flow chart of PhoCl evolution. The PhoCl variants discovered during the screening process are represented in blue rectangles. The final PhoCl2 variants are represented in green rectangles. The library generation method, specific mutations discovered, and other key points, are represented in white rectangles. 7

Photocleavable Proteins that Undergo Fast and Efficient …

DNA encoding the NES of MAPKK (NLVDLQKKLEELELDEQQ)26 and the NLS of SV40 (PKKKRKV)27were separately appended by PCR to the 5′ end of the gene encoding PhoCl. The gene encoding mCherry was inserted between the KpnI and XhoI sites of pcDNA3.1(+) (Thermo Fisher Scientific). Genes encoding NES … Visa mer Plasmid construction was performed through standard restriction-enzyme cloning. Pfu (Thermo Fisher Scientific), Taq (New England BioLabs), and Q5 (New England BioLabs) DNA polymerases were used for polymerase … Visa mer Overlap PCR was used to circularly permute the gene encoding mMaple, such that the original termini were joined with a GGSGG linker, and new … Visa mer All cell lines were grown from original frozen stocks without authentication or mycoplasma detection. The HEK cell line is listed in the International Cell Line Authentication … Visa mer The genes for mMaple, PhoCl, and PhoCl-MBP in pBAD/His B were used to transform E. coli strain DH10B or Top10. LB medium supplemented with 100 μg/mL ampicillin was inoculated with these transformants, and the … Visa mer WebbA non-protease-based optogenetic technique termed PhoCl (PhotoCleavable protein) based on violet-light-induced β-elimination reaction of a photoconvertible uorescent … daily provisions zoominfo

Adding Dynamic Biomolecule Signaling to Hydrogel Systems via …

WebbPhotocleavable Cadherin Inhibits Cell-to-Cell Mechanotransduction by Light. ACS Chem Biol, 2024 Genetically Encoded Photocleavable Linkers for Patterned Protein Release … WebbThe finalized PhoCl system has a broad absorbance from 400 to 500 nm, and the dissociation rate is 500 s. Many proteins, ≈ such as Cre recombinase, Gal4 transcription factor, Pannexin-1 ion channel, could be controlled by PhoCl.[10] WebbPhoCl may enable a generic approach for optogenetic activa- tion of enzymes if it is used as a photocleavable linker between an enzyme and its proteinaceous inhibitor. biomate corporation philippines photo