Prokaryotic expression and preparation of the polyclonal antibodies of E protein and domain Ⅲ in Zika virus / 中国人兽共患病学报
Chinese Journal of Zoonoses
; (12): 23-28, 2018.
Article
em Zh
| WPRIM
| ID: wpr-703062
Biblioteca responsável:
WPRO
ABSTRACT
To clone,express and purify the E Protein and EDⅢ of Zika virus in E.coli and prepare two kinds of polyclonal antibodies,the virus was amplified by Vero E6 cell culture.Total RNA was extracted by RT-PCR and reverse transcribed into cDNA.The prokaryotic expression vectors pET32a/E and pET28a/EDⅢ were constructed by cDNA sequence of E and EDⅢ gene.Then,recombinant plasmids were transformed into E.coli BL21 and induced by IPTG,and purified by Ni+ column affinity chromatography.BALB/C mice were immunized with purified recombinant proteins.Antiserum was collected and titer was determined by indirect ELISA.Western blot was used to detect the specificity.Results showed that the recombinant proteins were successfully expressed and purified.The titer of the polyclonal antibodies both reached 1:409 600.Western Blot analysis showed that the polyclonal antibodies could specifically recognize the recombinant proteins.Thus,the specific polyclonal antibody were successfully prepared,laying a foundation for further study on the pathogenesis,detection methods and immune strategies of Zika virus.
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Índice:
WPRIM
Idioma:
Zh
Revista:
Chinese Journal of Zoonoses
Ano de publicação:
2018
Tipo de documento:
Article