Cloning and expression of VP1 gene of Duck hepatitis virus genotype C and preparation of its polyclonal antibody

›› 2011, Vol. 23 ›› Issue (2): 0-262.

• 动物科学 •

Cloning and expression of VP1 gene of Duck hepatitis virus genotype C and preparation of its polyclonal antibody

ZHAO Li-na;CUI Yan-shun;REN Jian-ting;LI Jian-liang;HUANG Bing;LI Yu-feng;MA Xiu-li;*

1College of Animal Science & Veterinary Medicine, Shandong Agricultural University, Tai'an 271018, China; 2Institute of Poultry Science, Shandong Academy of Agricultural Sciences, Jinan 250023, China; 3Secondary Specialized School in Guangrao County, Shandong Province, Guangrao 257300, China

Received:1900-01-01 Revised:1900-01-01 Online:2011-03-25 Published:2011-03-25

Abstract

Abstract: The VP1 gene of strain JFX08 of Duck hepatitis virus（DHV） genotype C was amplified by reverse transcription-polymerase chain reaction (RT-PCR). To obtain expression plasmid pET-32a-VP1, the VP1 gene was cloned into pMD18-T and pET-32a(+)vectors. SDS-PAGE analysis revealed that the recombinant VP1 protein of genotype C of Duck hepatitis virus was expressed in Escherichia coli BL21 (DE3) at a high level after being induced with isopropylthio-β-D-galactoside (IPTG) stably. Western-blot revealed that the recombinant protein was recognized specifically by antisera against the genotype C of Duck hepatitis virus. Four-week-old SPF chickens were immunized with the purified recombinant protein. ELISA established by the purified recombinant protein and DHV genotype C revealed that the titer of antiserum was 1∶25 600 and 1∶51 200 respectively, which indicated that the recombinant protein had good immunogenicity．

Key words: Duck hepatitis virus, genotype C, VP1 gene, cloning and expression, polyclonal antibody

ZHAO Li-na;CUI Yan-shun;REN Jian-ting;LI Jian-liang;HUANG Bing;LI Yu-feng;MA Xiu-li;*. Cloning and expression of VP1 gene of Duck hepatitis virus genotype C and preparation of its polyclonal antibody[J]. , 2011, 23(2): 0-262.

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Cloning and expression of VP1 gene of Duck hepatitis virus genotype C and preparation of its polyclonal antibody

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