title: Universal and rapid method for purification of GFP-like proteins by the ethanol extraction.
abstract: GFP-like fluorescent proteins (FPs) are crucial in biological and biomedical studies. The majority of FP purification techniques either include multiple time-consuming chromatography steps with a low yield of the desired product or require prior protein modification (addition of special tags). In the present work, we propose an alternative ethanol extraction-based technique previously used for GFP purification and then modified for diverse FPs originated from different sources. The following recombinant FPs were expressed using Escherichia coli M15 (pREP4) strain as a host transformed with pQE30 plasmid bearing one of the target FP genes: TagCFP, TagGFP, TagYFP, TagRFP, TurboGFP, TurboRFP, Dendra2, TurboFP602 and KillerRed. Despite their diversity, all tested recombinant FPs were successfully purified and yielded a highly homogeneous product. The method is easily scalable for purification of any amount of protein and requires no expensive reagents and equipment.
authors.affiliation: Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Nauki Prosp. 6, 142290 Pushchino, Moscow Region, Russia.
authors.forename: Olga N
authors.initials: ON
authors.lastname: Samarkina
authors.forename: Anastasia G
authors.initials: AG
authors.lastname: Popova
authors.forename: Elena Yu
authors.initials: EY
authors.lastname: Gvozdik
authors.forename: Anna V
authors.initials: AV
authors.lastname: Chkalina
authors.forename: Ivan V
authors.initials: IV
authors.lastname: Zvyagin
authors.forename: Yulia V
authors.initials: YV
authors.lastname: Rylova
authors.forename: Natalia V
authors.initials: NV
authors.lastname: Rudenko
authors.forename: Konstantin A
authors.initials: KA
authors.lastname: Lusta
authors.forename: Ilya V
authors.initials: IV
authors.lastname: Kelmanson
authors.forename: Andrey Yu
authors.initials: AY
authors.lastname: Gorokhovatsky
authors.forename: Leonid M
authors.initials: LM
authors.lastname: Vinokurov
mesh_terms: D000818:Animals; D000431:Ethanol / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D000431:Ethanol