Universal and rapid method for purification of GFP-like proteins by the ethanol extraction.
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.
Samarkina
Olga N
ON
Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Nauki Prosp. 6, 142290 Pushchino, Moscow Region, Russia.
Popova
Anastasia G
AG
Gvozdik
Elena Yu
EY
Chkalina
Anna V
AV
Zvyagin
Ivan V
IV
Rylova
Yulia V
YV
Rudenko
Natalia V
NV
Lusta
Konstantin A
KA
Kelmanson
Ilya V
IV
Gorokhovatsky
Andrey Yu
AY
Vinokurov
Leonid M
LM
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
D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D000431:Ethanol