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Virologica Sinica, 18 (5) : 468-472, 2003
Research Article
Construction and Characterization of Recombinant CVI988 Expressing the Green Fluorescent Protein
Key Lab ofAnimal Diseases Diagnostic and Immunology,Ministry ofAgriculture,Nanjing Agriculture University,Nanjing 2 10095
The US2 gene nonessential for viral replication was amplified by polymerase chain
reaction(PCR)with DNA from CEF cells infected by MDV vaccine strain CV1988 as template,and was
cloned into T-easy vector;Th e expression cassette including GFP gene controlled by CM V promoter
and enhancer was cloned into US2 gene to give rise to a tran sfer vector pGUS2GFP,then the complex
of pGUS2GFP and DOTAP was transfected into CEF cells infected CVI988 strain virus.Recombinan t
rCVIGFP expressing the green fluorescence protein was selected an d purified.Th e characterization of
recombinant virus was evaluated n vivo and in vitro.Th e growth curve of recombinan t virus having
EGFP gene are similar to that of the parent viru s in CEF cells.Th e recombinat virus expressing GFP
could infect chicken and be reisolated from the chickens following intra-abdominal inoculation of
  Published online: 5 Oct 2003
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