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采用q PCR与病毒滴度测定观察RNA干扰(RNA interference,RNAi)对大鲵蛙病毒(Chinese giant salamander ranavirus,CGSRV)主要衣壳蛋白(major capsid protein,MCP)、甲基转移酶(methyltransferases,MTases)、DNA多聚酶(DNA polymerase)基因表达与病毒增殖的影响。结果表明,小干扰RNA(small interfering RNA,siRNA)能推迟鲤鱼上皮瘤细胞(epithelioma papillosum cyprini,EPC)出现病变,且病变程度也较对照组轻。在各功能基因表达量上,NC-FAM对照组与阴性对照组差异不显著(P>0.05);而干扰组的干扰率极显著高于阴性对照组,其中siR-DP-1组siRNA对MCP基因的干扰率为79%,极显著高于其余组(P<0.01);siR-MT-1、siR-MT-2组siRNA对MTases基因的干扰率为77%,极显著高于其余组(P<0.01);siR-DP-1组siRNA对DNA polymerase基因的干扰率为79%,极显著高于其余组(P<0.01)。干扰实验组病毒滴度与NC-FAM对照组和阴性对照组相比也有不同程度的降低。阴性对照组lg TCID50为8.362,NC-FAM对照组lg TCID50为7.848,siR-MCP、siR-MT、siR-DP实验组最低lg TCID50分别为5.764、5.317、5.362。证实RNAi能够抑制CGSRV主要功能基因的表达并影响病毒的复制。
The effects of RNA interference (RNAi) on the expression of major capsid protein (MCP), methyltransferases (MTases) in Chinese giant salamander ranavirus (CGSRV) , DNA polymerase gene expression and virus proliferation. The results showed that small interfering RNA (siRNA) could delay the pathological changes of epithelioma papillosum cyprin (EPC), and the degree of lesion was also lighter than that of the control group. There was no significant difference between the NC-FAM control group and the negative control group (P> 0.05), while the interference rate in the interference group was significantly higher than that in the negative control group The gene interference rate was 79%, which was significantly higher than the other groups (P <0.01). The siRNA of siR-MT-1 and siR-MT-2 had a 77% P <0.01). The interference rate of siR-DP-1 siRNA to DNA polymerase gene was 79%, which was significantly higher than the other groups (P <0.01). Interference in experimental group virus titers and NC-FAM control group and negative control group compared to varying degrees of reduction. Negative control group lg TCID50 8.362, NC-FAM control group lg TCID50 7.848, siR-MCP, siR-MT, siR-DP experimental group lg TCID50 were 5.764,5.317,5.362. Confirmed that RNAi can inhibit the expression of CGSRV major functional genes and affect the virus replication.