Abstract: To obtain the major structural protein hexon protein of serum type 4 avian adenovirus by prokaryotic expression; A pair of specific primers was designed according to the gene sequence of the hexon protein of Fowl adenovirus serotype 4 (FAdV-4) in GenBank. The full-length sequence of hexon gene was amplified by ordinary PCR. The serotype 4 avian adenovirus gene fragment obtained by PCR amplification was inserted into the vector pMD18T after inserting restriction enzyme sites at both ends of the gene. The PCR analysis and sequencing analysis showed that the gene was verified with the prokaryotic expression vector pET-32a. The constructed expression vector was transformed into BL21(DE3) and induced to express with 1 mmol/L IPTG. Analyzing the recombinant protein by SDS-PAGE and Western Blot; By the double-enzyme digestion and sequence analysis,the prokaryotic expression vector of pET-32a-hexon was successfully constructed. The size of the inserted hexon protein gene fragment was 2 814 bp. The recombinant protein expression was highest at 3 h after induction with 1 mmol/L IPTG,and the recombinant protein was mainly inclusion body forms exist in sediments. The SDS-PAGE analysis showed that the fusion protein had a molecular weight of 118 ku. Western Blot analysis showed that the fusion protein could be specifically recognized by anti-FAdV-4 serum and HIS tag antibody. This experiment successfully expressed the major structural protein hexon protein of serum type 4 avian adenovirus,and the fusion protein obtained had good reactogenicity,which could be used in the further research of serum type 4 avian adenovirus. It laid a good foundation for the prevention and treatment of serum type 4 fowl adenovirus disease.

Key words: Fowl adenovirus serotype 4(FAdV-4), Hexon protein, Prokaryotic expression

CLC Number: 

• Q78

• S432.4