Soluble expression and purification of VP1 protein from O-type foot-and-mouth disease virus in Escherichia coli and its nanometer structure observation

doi:10.16098/j.issn.0529-1356.2018.01.020

Abstract

Abstract:

Objective To establish a method to express the soluble VP1 protein from foot-and-mouth disease virus type O in Escherichia coli, (E. coli) and to observe its nanoparticles. Methods VP1 gene from FMDV O/MYA/7/98 strain was obtained according to the sequence of O-type foot-and-mouth disease virus. Truncation and optimization were performed. The fusion tages were screened to obtain soluble expression of SeFnt16798 protein. The 73 aa ferritin protein was isolated from Salmonella enteric and linked with the VP1 protein, which was named SeFnt16798. Prokaryotic expression vectors were fused with nine different fusion tages(Grifin, GST, MBP, Sumo, Thioredoxin, γ-crystallin, ArsC, PpiB and CeHSP17)fragments were constructed. These recombinant plasmids were transformed into Escherichia coli BL21 (DE3) and induced by isopropyl β-D-thiogalactiside(IPTG). SDS-PAGE analysis was used to identify the soluble expression of recombinant protein after ultrasonic decomposition. The fusion tag that could significantly promote soluble expression of SeFnt16798 protein was selected. Abundantly inducible expression recombinant protein was purified by Ni-NTA purification system and detected by electron microscopy. Results Recombinant expression of SeFnt16798 was successfully constructed and had the high purity MBP-SeFnt16798. The result of electron microscopy showed that MBP-SeFnt16798 formed nano-sized particles. Conclusion We established a stable method to obtain recombinant protein of SeFnt16798, which may lay a foundation for the development and subsequent study of FMDV structure vaccine.

Key words: O-type foot and mouth disease virus, VP1 protein, Ferritin, Fusion tag, Nanoparticle, Electron rnicroscopy, Escherichia coli

GUO Yu-kun MING Sheng-li GUO Wan-ying YANG Guo-yu GUO Yu-jie. Soluble expression and purification of VP1 protein from O-type foot-and-mouth disease virus in Escherichia coli and its nanometer structure observation[J]. Acta Anatomica Sinica, 2018, 49(1): 118-123.

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