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| Bioactivity of the Escherichia coli-derived Norovirus P Particles and Its Shape Under TEM |
| YANG Gui-hua, CHEN Lu-lu, WANG Xiao-lei, LI Shan-shuang, ZHAO Ling-xia |
| Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China |
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Abstract Human noroviruses(NVs)are primary causes for epidemic nonbacterial gastroenteritis in the world, and high-risk for young children ,elderly peoples and crowd in public location. Currently, usage of the vaccine is a major strategy to prevent infection of NVs and outbreak of acute gastroenteritis in clinic. In order to develop a new vaccine, a PGENE(1 002bp), which encoding P particle of the NVs capsid protein(strain VA387) was designed and synthesized according to tomato codon usage bias, and a DNA sequences encoding 6×His-tag was placed at 5' end of the PGENE for easy to purify the recombinant P particle. Subsequently, the prokaryotic expression vector of the pET32a-PGENE was created and introduced into strain BL21 (Escherichia coli). The BL21 cells harboring pET32a-PGENE were induced by IPTG (isopropyl β-D-1-thiogalactopyranoside) in 1mmol/L final concentration to express the recombinant P particles at 16 ℃. And then, the recombinant P particles were purified by Ni-NTA resin and confirmed by western blotting technique.The E.coli-derived P protein was successfully assembled into global shape and found under TEM (transmission electron microscopy). The antiserum which is from the immunized BALB/c mice using P protein can be specifically binding to the P particles. These results will lay a foundation to produce oral plant vaccine using tomato fruits.
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Received: 12 November 2012
Published: 25 March 2013
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