Construction of prokaryotic expression vector for F protein epitope gene cascade of Newcastle disease virus.

Three epitope configuration regions of Newcastle disease virus F gene from SPF chicken embryo about 81,108,105 bp were cloned by RT-PCR using three pair of primers.The tandem-arranged multiple epitope gene of the F gene about 306 bp was obtained by appending complementary endonuclease of genetic engineering means.The fragment of Newcastle disease virus F gene was cloned into pET-32-a.The recombinant plasmid,namely pET-32-a-F306,was transformed into Escherichia coli BL21 and His-F306 fusion protein was induced to express,A MW of the fusion protein was about 31 000 as analyzed by SDS-PAGE,the partial segment size in F gene was 11 000.The purified fusion protein was used to immunized mice and the specific antibody was observed,which was identified further in immunodifusion,ELISA and Westtern-blot.The results indicated that this expressed fusion protein had a favorable antigenieity.