Expression,Purification and Crystallization of Severe Fever with Thrombocytopenia Syndrome Virus Envelope Glycoprotein Gc
To understand the mechanism of virus assembly and adhesion to host cells,this study pre-pared glycoprotein Gc of severe fever with thrombocytopenia syndrome virus(SFTSV)by insect cells ex-pression system,and provides an important theoretical basis for further antiviral drug development.The se-cretory expressive construct of recombined Gc ectodomain protein was designed.The purified protein was expressed by insect cells expression system.The results of SDS-PAGE analysis after purified by nickel af-finity chromatography and size-exclusive chromatography shows the high purity of Gc ectodomain protein.Crystal screening was carried out by hanging drop vapor diffusion method at a constant temperature of 16 ℃.After optimization of the pH and precipitant concentration,crystals were grown under two crystalli-zation conditions:2 mol/L ammonium sulfate;0.1 mol/L Hepes,pH 7.5,0.8 mol/L Sodium phosphate monobasic monohydrate,and 0.8 mol/L potassium dihydrogen phosphate.The Results showed that the Gc ectodomain protein with high purity was obtained.The protein crystals were grown.
severe fever with thrombocytopenia syndrome virusinsect cells expression systemcrystallizationglycoprotein Gc
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