Molecular Biological Characterization Of Avian Reticuloendotheliosis Virus

The avian reticuloendotheliosis virus proviral genome cDNA clone (pPB101) was transfected into chicken embryo fibroblast (CEF) cell by mixed with lipofectine. Using the monoclonal antibody to REV and the anti-sera against the REV env gp90-GST fusion protein. The molecular cloned virus was detected by IFA.We also amplified the gp90 from the cells infected with the molecular cloned virus by polymerase chain reaction. All these results indicated the recombinant plasmid containing the total REV genome cDNA is infectious.The proviral genome cDNA clone of SNV strain was digested, and the fragments was cloned into the pUC18 vector, sequenced, finally we got the sequence of SNV genome.The two pairs of primers were designed and synthesized according to the SNV strain env sequence. The env genes of the REV isolates isolated from China were amplified by polymerase chain reaction (PCR). The PCR products were cloned into the pUCm-T vector. After being sequenced, the two env genes were compared with that of SNV strain. The results showed that the two isolates have more than 95% homology with the SNV strain.The env gp90 was amplified from the SNV strain by PCR and was cloned into the downstream of GST gene in pGEX-6P-l vector according to the correct open reading frame, and this recombinant was transformed into BL21 for expression. The 64KD fusion protein was identified by SDS-PAGE.