Expression and Identification of VP1 Gene of Duck Hepatitis A Virus Type Ⅰ in Insect Cells
GU Lingling
WU Meng
ZHU Shanyuan
WANG Anping
Abstract：In order to express the main structural protein VP1 of duck hepatitis A virus type Ⅰ (DHAV-Ⅰ)in insect cells,one pair of specific primers were designed according to the published genome sequences of DHAV-Ⅰ to amplify VP1 gene by RT-PCR,the amplified fragment was cloned into baculovirus expression vector pFastBac1.The recombinant vector pFB-VP1 was transformed into E.coli DH10Bac competent cells,and the positive recombinant bacmid rBacmid-VP1 was screened according to the resistant and the blue-white plague screening,confirmed by PCR.The recombinant bacmid rBacmid-VP1 was transfected into the Sf 9 insect cells by liposome to obtain the recombinant baculovirus,and was named as rBac-VP1.The result of Western blot showed that the molecular weight of the recombinant proteins was about 27 ku,and could be recognized by the antiserum of DHAV-Ⅰ VP1.Indirect immunofluorescence analysis proved that the recombinant proteins could be recognized by the positive anti-virus serum.These results suggested that the main structural protein VP1 of DHAV-Ⅰ with immunogenicity was expressed successfully in insect cells.
Keywords：Duck hepatitis A virus type ⅠVP1 geneInsect cellsExpressionIdentification
Publication Date：2018-01-01
Online Publishing Date：2026-09-12(First online date of this platform, not the publication date of the document)
Pages：4( 114-117 )