Expression and purification of enterovirus type 71 polyprotein P1 using Pichia pastoris system.
Enterovirus type 71(EV71) causes severe hand-foot-and-mouth disease (HFMD) resulting in hundreds of deaths of children every year; However, currently, there is no effective treatment for EV71. In this study, the EV71 poly-protein (EV71-P1 protein) gene was processed and cloned into the eukaryotic expression vector pPIC9k and then expressed in Pichia pastoris strain GS115. The EV71 P1 protein with a molecular weight of 100 kD was produced and secreted into the medium. The soluble EV71 P1 protein was purified by column chromatography with a recovery efficiency of 70%. The result of the immunological analysis showed that the EV71 P1 protein had excellent immunogenicity and could stimulate the production of EV71-VP1 IgG antibody in injected rabbits. We suggest that EV71-P1 protein is an ideal candidate for an EV71 vaccine to prevent EV71 infection.
College of life science, Jianghan University, Wuhan, 430056, Hubei, China. hanxue_whdx@163.com
Xue
X
Han
Xiaoling
X
Ying
Hao
H
Huang
Shili
S
Zhou
Qi
Q
Huang
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D003470:Culture Media / Q000737:chemistry; D004770:Enterovirus / Q000235:genetics*; D015870:Gene Expression; D008970:Molecular Weight; D010843:Pichia / Q000235:genetics*; D020815:Polyproteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D014764:Viral Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D014765:Viral Vaccines / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
D000914:Antibodies, Viral; D003470:Culture Media; D020815:Polyproteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D014765:Viral Vaccines