Construction Of Recombinant Fowl Adenovirus Type Ⅰ Expressing IBDV VP2 Protein (rFAVI-VP2)

Fowl Adenoviruses have a wide host-cell range, most of which are nonpathetic and could be propagated in large amount easily. HE Xiu-Miao previously isolated a strain of Fowl Adenovirus group I and constructed universal transfer vector pFAVI-CMV. Infectious bursal disease virus (IBDV) VP2 gene from pcDNA3.1-VP2 was subcloned into pFAVI-CMV by EcoRâ… and Nheâ… double digestion. The recombinant transfer vector was designated as pFAVI-VP2. Restriction endonuclease digestion identified it right. In the presence of Lipofectin, recombinant virus rFAVI-VP2 was obtained through homogeneous recombination between wild Fowl Adenovirus (ws-FAVI-JS) and pFAVI-VP2 DNA in CEK. Indirected Immunofluorescence assay (IFA) was used to confirm the recombination and expression of IBDV-VP2 protein. The recombinant virus was purified through serial dilution of the supernant and double layers Agarose covery methods. Finally we got a strain of purified recombinant virus (rFAVI-VP2). The recombinant virus was propagated in large stocks by inoculating 9 days SPF chicken embryo. PCR and ELISA methods were employed to detect the amplification of 13^th. passage recombinant virus and both showed the virus could be passed stabliy in vitro.