Effect of Host Protein Rab11a on Replication of Bovine Viral Diarrhea Virus
To investigate the effect of host protein Rab11a on the proliferation of Bovine viral diarrhea virus(BVDV),the MDBK cell line with stable Rab11a gene knockout was constructed by CRISPR/Cas9 gene editing technology and the guide RNA expression vector targeting Rab11a gene was designed in the present study.The protein expression of Rab11a gene knockout was examined by Western blot.The levels of viral RNA and double strand RNA,virus titer and CPE were tested in qPCR and immunofluorescence assay.The results showed that Rab11a KO cells with the gene knockout were constructed as demonstrated in Western blot and Rab11a knockdown significantly inhibited the accumulation of BVDV 5'UTR mRNA and dsRNA levels.A large number of cytopathic changes were observed in the Scramble cells at 36 h post BVDV infection,such as shrinkage,fragmentation,vacuolation and fall off,etc.The CPE development was significantly lower in Rab11a KO cells than Scramble cells.In addition,the BVDV titer in Rab11a KO cells was significantly lower than that of Scramble cells at 12 h post infection.In conclusion,knockout of Rab11a gene in MDBK cells inhibited the intracellular replication of BVDV.The interaction between host protein Rab11a and BVDV was more understood from the discovery in this study.
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