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

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. Key words: Rotavirus; LLR; VP8* Protein