The Expression Divergence Of Mx Gene In SPF Shao Duck Embryonated Eggs Infected By DHAV-1 And DHAV-3

Interferons?IFNs?are the first line of defense against viral infections in vertebrates.Mx has high value for investigation since it is the indicator of I-IFN.There are few study about duck Mx.This study showed the relationship of mRNA level of Mx and IFN-? and DHAV-1/-3 after injected with DHAV-1/-3 in duck embryonated eggs.This study will provide a theoretical basis to investigate the immune response against virus infection.We designed a specific pair of primer to amplify the CDs of Shao duck Mx gene.The cDNA of Shao duck? s kidney was used as template to perform PCR procedure to amplify Mx.Then subcloned1435-2148 bp of CDs to prokaryotic expression vector pET-30a?+?,expressed in E.coli RosettaTM?DE3?pLysS with the molecular mass 35 kDa.The rDupET-30a-Mx was purified by Ni-NTA column.The PAb against duck Mx protein was prepared with rDup ET-30a-Mx.The titer of PAb was above 1:102400 detected by I-ELISA.Western blot showed that the PAb can recognize Mx protein in adult duck and duck embryonated eggs.10³ELD₅₀ DHAV-1 or DHAV-3 were injected to 15 days SPF duck embryonated eggs.The heart,liver,stomach,intestine and muscle were collected after injected 3h,6h,12 h,24h,48 h.The relative m RNA levels of Mx,IFN-? and virus load in those tissues were detected by qPCR.The Mx protein were detected by Western blot.The results showed that the mRNA of Mx and IFN-? were parallel during 3h to 12 h in those five tissues;the virus load was declined as Mx was increased in heart,liver and stomach.