To evaluate canine adenovirus type 1 vaccine strain (Cannaught Laboratory Limited, CLL) as recombinant vaccine and gene transfer vector.Recombinant virus CLLEGFP which contains enhanced green fluorescent protein(EGFP) reporter gene was constructed. CLLEGFP was used to infect various human derived cell lines(293, Hela, CO, SW, Hep-2 and CAM) by inoculating intraperitoneally(i.p.), intravenously(i.v.) and intramuscularly (i.m.) to Kunming mice other than oral administration. Various tissue samples of the mice were collected at multi-time point for observing EGFP green fluorescence. Anti-EGFP antibodies were detected by Western blot analysis in the sera after 4 weeks.CLLEGFP can infect various human derived cell lines and express EGFP. EGFP green fluorescence were observed in liver tissue cells after i.p. transducing 3 days. All immune inoculation ways above could induce Kunming mice producing anti-EGFP antibodies which were identified by Western blot analysis.These results indicate that CLL possess powerful potential as recombinant vaccine and gene transfer vector.