Purification and influencing factors of myelin regulatory factors by nickel metal chelation
To study the nickel metal chelation chromatography purification method and influencing factors of myelin gene regulatory factor(MYRF),BL21 E.coli with MYRF gene was used as the host bacterium.Different concentration gradients of imidazole and EDTA were set for nickel metal chelation chromatography column purification.The study was analyzed by protein concentration measurement,coomassie brilliant blue staining,and SDS-PAGE analysis.The effect of binding time between protein supernatant and nickel filler on MYRF protein purification were further studied.The results showed that MYRF concentration obtained by imidazole was proportional to the elution solution concentration,and a 0.2 mol·L-1 imidazole solution could obtain MYRF protein with high purity.Although low concentration(0.05 mol·L-1)EDTA can quickly ob-tain high concentration MYRF protein,the content of impurities was also high.Meanwhile,there was a positive correla-tion between MYRF concentration and binding time.In summary,MYRF protein nickel metal chelation chromatography is influenced by multiple factors such as the type of eluent and the binding time of fillers,providing reference basis for the selection and optimization of MYRF protein purification strategies.
nickel columnimidazoleEDTAprotein purificationbinding time
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