| Innate immunity is a series of natural protective ability that set up by our body in the process of the development for a long-term, which provide a barrier can quickly hinder the spread of the virus infection. RING finger protein (RNF11) is one of the family members of the RING finger family, also the family members of E3 ubiquitin ligase, which plays an important role in the antiviral pathway. However, there has been no report about whether is involved in the innate immunity of chicken antiviral pathway and in the regulation of the inflammatory response. In this study, the full-length CDS coding sequences of chicken RNF11 genes were cloned firstly, analysis the information of the sequence and predict its related features of Protein. Then,1 day-old SPF chicken was Injected subcutaneously ALV-J, and the differential expression of RNFll, IFN-a and IFN-β genes were determined by real-time PCR to research the differential expression of RNF11 gene at 14,21,28 and 35 days in the three tissue:Bursa of fabricius, spleen and thymus. Finally, we construct the expression vector and the interference carrier of RNFll gene, the transfect them into chicken’s fibroblast cells. We analyzes the expression of IFN-β gene influenced by RNF11 gene at 24 and 28 hours after transfection (HPS),to further validate the effect of RNF11 gene on antiviral pathway, Besides, to detect the expression of the inflammatory factor IL2 and IL10 gene,and to discussion the effect of RNF11 gene on inflammatory response. The results are listed as follows:1) We obtained a 465 bp sequence which has a stable structure and encoding 154AA after analysis the CDS sequence of RNF11 gene. We predict it as a transmembrane and hydrophobicity protein, which molecular weight is 17.4156kDa. The protein has nine phosphorylation sites and is one of the family members of the RING finger family, which may play a role in the nucleus and cytoplasm.2) After 1 day-old SPF chicken was injected subcutaneously ALV-J, the highest expression of ALV-J gene was at 21 day in the three tissue, the most copies of viral replication, and the trend of the four time points in 14,21,28 and 35 days are basically identical of the expression of ALV-J gene. In Bursa of fabricius, ALV-J causes the expression of RNFll gene extremely significant (p<0.01) increased in each time point, whereas the expression of IFN-a and IFN-β gene were significant (p<0.05) or very significant (p<0.01) increased, in the spleen, the expression of RNF11 gene is very significant (p<0.01) increased at 14 and 21 day, there is no significant difference in 35 day, in addition to the expression of IFN-a and IFN-β gene no significant difference in 21 day, and the expression of IFN-a and IFN-β gene were significantly or extremely significantly higher than the control group. In thymus tissues, ALV-J also causes the expression of RNF11 gene is significantly or extremely significantly higher than control group at 14,21 and 28 day. The expression of IFN-β gene is differences in 35 days and no significant contrast. It is indicate that ALV-J causes the rise expression of interferon gene in the body, caused the body’s antiviral response, the expression of RNF11 gene is raised, suggest that RNF11 gene plays a role in chicken antiviral pathway.3) Next, the chick embryo fibroblast(CEFs) were infected with the vector of overexpress and interference to testify ulteriorly the role of KNF11 gene on antiviral pathway. The result showed that the expression of IFN-β gene was extremely significantly lower than control at 24 hour or 28hour post-infection when we overexpress the RNF11 gene. But, the expression of IFN-β gene was extremely significantly higher than control after interfed the RNF11 gene.4) By expressing and interfering the expression of RNF11 gene in chicken fibroblast cells,we found that the expression of IL2 and IL10 gene were increased after overexpress RNF11 gene, but the expression of IL2 and IL10 gene were dropped significantly when interfering the expression of RNF11 gene, which reveals RNF11 genes play an important role in the inflammatory response. |
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