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Tuning the Flexibility of Glycine-Serine Linkers To Allow Rational Design of Multidomain Proteins.

Biochemistry. 2017-12; 
van RosmalenMartijn, KromMike, MerkxMaa
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PCR Cloning and Subcloning … the linker sequences (see Figure S1 for sequence details). pET-28a(+)-CLY-(G 2 S 4 ) 9 , pET-28a(+)-CLY-(GS 5 ) 9 , and pUC-57-(S 6 ) 9 were ordered from Genscript (Piscataway, NJ). To obtain pET-28a(+)-CLY-(S 6 ) 9 , the … Get A Quote

摘要

Flexible polypeptide linkers composed of glycine and serine are important components of engineered multidomain proteins. We have previously shown that the conformational properties of Gly-Gly-Ser repeat linkers can be quantitatively understood by comparing experimentally determined F?rster resonance energy transfer (FRET) efficiencies of ECFP-linker-EYFP proteins to theoretical FRET efficiencies calculated using wormlike chain and Gaussian chain models. Here we extend this analysis to include linkers with different glycine contents. We determined the FRET efficiencies of ECFP-linker-EYFP proteins with linkers ranging in length from 25 to 73 amino acids and with glycine contents of 33.3% (GSSGSS), 16.7% (GSSS... More

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