[Optimizing expression of the capsid protein VP2 from human Bocavirus and establish it's seroepidemiology assying methord].
2012
Objective To obtain sufficient recombinant VP2 protein of human Bocavirus and establish it's seroepidemiology assying methord.Methord The capsid protein VP2 DNA genes of HBoV1 and 2 were optimized in accordance with the usage of the favorite codons in E.coil so as to enhance its protein expression in prokaryotic expressing system.The protein was purified by Ni-NTA column,and its antigenicity was determined by Western Blot.Then establish ELISA to detect the specific anti-VP2 IgG antibodies against HBoV1 and 2 in healthy children aged 3 - 6 years in Nanjing,China.Results The recombinant protein 6 × His-VP2 was produced in a larger quantity at 25℃ induced by IPTG (lmmol/L) over night and purified by Ni-NTA column.Seropositive rates of HBoV1and 2 were 62.2% and 55.5% and their mixed seropositivity was 37%.Conclusion The optimizing expression of the capsid protein VP2 from human Bocavirus constructed successfully and get a high yield under certain conditions.The established ELISA could be used to further analyze seroepidemiology of HBoV in china.
Key words:
Human Bocavirus; Codon; Expressied sequence tags; Epidemiology, melecular; Enzyme-linked immunosorbent assay
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