Cloning And Prokaryotic Expression Of PRRSV GP4 And GP5 Protein Gene

Porcine reproductive and respiratory syndrome (PRRS), a hyperinfection disease caused by PRRS virus, which is mainly characterized on premature birth gravid sow, miscarriage, fetal death, mummy fetus, week fetus and respiratory passage disease in piglet.A new strain of high pathogenicity PRRS was isolated and identified from the swine infected with PRRSV in Baicheng Area. This virus was isolated from the lung tissue of sow that can infected with MARC-145 cells and was inspected by RT-PCR, physical and chemical characterization, serological tests, electron microscope or other methods, then named BC strains.In this research, according to the complete nucleotide sequence of PRRSV in GeneBank, two pair of primers were designed to amplify ORF4 and ORF5 gene of the BC strain. Following Sequencing, the length of BC strain GP4 gene was 537bp, encoding 179 amino acids;but the length of BC strain GP5 gene was 603bp, encoding 200 amino acids. This two kinds of major structural protein genes of BC strain were cloned in prokaryotic expression vector respectively, WeStern-blotting showed that the recombinant protein could react with the porcine polyclonal antibody against PRRSV, the indirect ELISA to detect PRRSV was established, which will provide a good fundament for development of PRRSV antibody surveillance kit.