Expression Of E Gene Of IBV And Analysis On The E Genes Of Different IBV And Different Coronaviruss

Avain Infectious bronchitis (IB) is caused by the infectious bronchitis virus (IBV) which belongs to the Coronavirus. It is an eveloped positive-strand RNA viruse, also the biggest one in the Coronavirus. This virus has four proteins, which are sipke protein (S), matrix protein (M), necleocapsid protein (N) and the envelope protein (E). We used do much more study on the proteins of S, M and N than on the protein E.The third ORF of IBV encodes the envelope protein. Its molecular weight is 12.4KD. It can company with the M protein to formate Virus-like particle (VLP), which plays an important role in the mucosa immunity. In the process of reproduce, E protein takes part in the formation of VLP and budding of the whole virus. Recently, we found there is a lot of E protein in the late period of the IBV infection, and presumed the E protein is related to the apoptosis.In this study the envelope protein (E) gene of Avian Infectious Bronchitis Virus (IBV) M41 strain was cloned, and subcloned into prokaryotic expressing vector pGEX-6P-1. Fusion protein was induced by IPTG after being verified. The GST protein expressed by the pGEX-6P-l+BL21 was purified, the GST protein was immuned into mouses according to the normal immunity process, and the multi-cloned serum was obtained. The multi-cloned serum was used to test the fusion protein GST-E and its activity, in addition to this, the GST-E was used to test the antibody in the blood of SPF chickens which has been infected by the IBV, all these were done to explore the expression of the E protein in the Escherichia coli and the expectation of the E protein used in the examination of the virus. Meanwhile, the E protein sequences in many strains of IBV and this of other types of corovavirus were compared, in order to offer some information to the future study of the E.The M41 E protein gene was cloned into the pGEX-6P-1, and the GST-E fusion protein was got, which was about 38.4 KD. In the western-blotting test, there was a reaction lane in the area of 38.4 KD, showed that the GST-E had reaction activity. Western-blotting was also used to identified the blood of SPF chickens which were infected by IBV, found the anti-E antibody can be found ever since the 5th day until the 14^th day after infected. The IBV E sequences comparison showed that although the blood types were different, the sequences of the E protein were conservative. But the E genes of IBV and TCoV had low homology with other types of corovavirus', while the sequences of different types of corovavirus were being analysed.