Optimizing expression of the capsid protein VP2 from human Bocavirus and establish it's seroepidemiology assying methord.
- Author:
Ye-Xia HAO
1
;
Ji-Min GAO
;
Yu JIN
;
Xiao-Le LI
;
Jing-Song LI
;
Zhi-Ping XIE
;
Yuan-Yun AO
;
Xi-Qian CHEN
;
Ke-Na CHEN
;
Zhao-Jun DUAN
Author Information
- Publication Type:Journal Article
- MeSH: Capsid Proteins; analysis; genetics; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Female; Human bocavirus; isolation & purification; Humans; Male; Polymerase Chain Reaction; Seroepidemiologic Studies
- From: Chinese Journal of Experimental and Clinical Virology 2012;26(1):18-21
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo obtain sufficient recombinant VP2 protein of human Bocavirus and establish it's seroepidemiology assying metbord. METHORD: Tbe capsid protein VP2 DNA genes of HBoV1 and 2 were optimized in accordance with tbe usage of the favorite codons in K 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.
RESULTSThe recombinant protein 6 x His-VP2 was produced in a larger quantity at 25 degrees C induced by IPTG (1 mmol/L) over night and purified by Ni-NTA column. Seropositive rates of HBoV1 and 2 were 62.2% and 55.5% and their mixed seropositivity was 37%.
CONCLUSIONThe 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.