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Purification scheme. Purification scheme. (A) Line drawing of CelTag fragment from CelTag plasmid. PCR (polymerase chain reaction) of fragment was performed using primers with 50 nucleotides of sequence identity to the site of chromosomal insertion and 21 nucleotides of complementarity to the CelTag. The sequences of fragment and primers used in this study are found in File S1 and Table S1, respectively. (B) Schematic purification scheme. Pgk1 with C-terminal CelTag binds efficiently to cellulose surface and elutes specifically with TEV (Tobacco Etch Virus) protease. Elution with SDS (sodium dodecyl sulfate) results in low, variable levels of contamination from nonspecifically bound proteins. CBM3, family 3 cellulose binding module. Brian H. Carrick et al. G3 2015;6: ©2016 by Genetics Society of America
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