Expression and purification of the VP8 * protein of group A rotavirus vaccine strain LLR

VernacularTitle:A组轮状病毒疫苗株LLR VP8*基因的原核表达及纯化
Author: Nijun GUO ¹ ; Xiaoman SUN ; Dandi LI ; Youde CAO ; Zhaojun DUAN
Author Information

1. 湖南师范大学第一附属医院湖南省人民医院检验科
Keywords: Rotavirus; LLR; VP8 * Protein
From: Chinese Journal of Experimental and Clinical Virology 2015;29(5):452-454
CountryChina
Language:Chinese
Abstract: Objective To express the VP8 * protein of the rotavirus vaccine strain LLR in the E.coli with the pGEX4T-1 vector,which is important for the further research of the VP8 * protein functions.Methods The LLR VP8 * gene was obtained by virus RNA extraction and RT-PCR.Then it was cloned in the expression vectors pGEX4T-1.The recombinant plasmid pGEX4T-1-VP8 * was transformed to the E.coli BL21.Then the VP8 *-GST fusion protein was expressed and purified by the affinity chromatograph.The protein of interest was validated by SDS-PAGE and Western Blot.Results The molecular weight of the VP8 *-GST fusion protein was about 52 000 according to the SDS-PAGE.The bands of both 52 000 and 26 000 were shown in the Western Blot with the antibody against GST.Conclusions The LLR VP8 * gene was obtained and cloned to the pGEX4T-1 vector.Moreover,the solvable VP8 *-GST fusion protein was successfully expressed and purified.