In order to investigate the targeting relationship of miR-130c-5p to the potential target gene g in snake-head vesiculovirus(SHVV)infection and its effect on viral replication,the changes of viral gene and protein levels and miR-130c-5p in SHVV were determined in this study by quantitative real-time PCR(qRT-PCR)and Western blot techniques using channel catfish ovary(CCO)as experimental materials.In addition,the target sequence cor-responding to miR-130c-5p on the g gene of SHVV was cloned into the plasmid pmirGLO,and the plasmid pmir-GLO-G was constructed for dual luciferase reporter assay for target gene verification.The results showed that the expression levels of miR-130c-5p and g genes were significantly up-regulated with the increasing time and dose of SHVV infection.Further experiments showed that co-transfection of miR-130c-5p mimic and pmirGLO-G plas-mid significantly inhibited luciferase activity,while transfection of miR-130c-5p inhibitor significantly up-regu-lated the fluorescence signal of pmirGLO-G reporter vector.In addition,overexpression of miR-130c-5p signific-antly reduced the mRNA and protein expression of the viral g gene,while inhibition of miR-130c-5p up-regulated the mRNA and protein expression levels of the g gene.The results showed that miR-130c-5p inhibited the prolifer-ation of SHVV by targeting the g gene of SHVV and causing the degradation of G protein.The results of this study provide an important basis for understanding the pathogenic mechanism of microRNA regulation of SHVVV,and provide theoretical support for the development of anti-SHVV vaccines and other drugs.
维普
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