Cloning, Expression And Application Of Structural Protein Gene Of Avian Infectious Bronchitis Virus

Avian Infectious Bronchitis (IB) is a highly acute and contagious disease of chikens caused by Avian Infectious Bronchitis virus (EBV), and has been one of the virus diseases which highest hazard the fowl industry. IBV is lack of haemagglutination speciality and hence the haemagglutination inhibition (HI) test couldn't be employed for serotyping and detecting the Serological antibody against IBV after the chikens infected of IBV or vaccinated. But detecting the IBV Serological antibody has the flowing usages. 1 It provides the foundation for appropriate vaccinated time.2.It can be used to value the vaccinated effect.3. It also can be assistant identification of EBV.It has reported tracheal organ culture (TOC) had been used to detect IBV antibody, but Virus neutralisation is laborious, time cosuming and impractical, hard to popularize. The enzyme-linked immunosorbent assays (ELISA) which is widely used to identify IBV-infected flocks (broilers) based on high antibody titres i s very sensitive, objective and cost. At present Commercial kits for ELISAs are available - these are based on several different strategies for the detection of IBV antibodie. Such tests using the whole IBV as coated antigen for detecting the antibody against IBV need culture much of IBV, and is easy to spread IBV. Our previous studies demonstrated that the IBV was easy broken, and the virus coronal was easy lost When IBV was isolated from the allantoic fluids of eggs, so It was hard to harvest the whole IBV antigen. In this study, in order to provide the foundation for detection of the IBV antibody, the structural genes of IBV were amplified and expressed based on the reported gene sequences, and The affinity-purified expressed protein was used as a coating antigen for an ELISA for serum and tears antibody detection of the chicken of infected by IBV M41.In this study, Nucleocapsid protein (N protein) Spike Protein (S protein) and partial S1 gene fragment (n.t.1144-1611) were cloned and expressed .The main results of our study are as follows:1. The N gene, S gene and partial S1 gene fragment (n.t.1144-1611) were cloned, sequenced and compared with those related genes collected in GenBank.Using...