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国家科技重大专项(2011ZX08006-001)

作品数:2 被引量:1H指数:1
相关作者:王铁东逄大欣戴祯王侃侃欧阳红生更多>>
相关机构:军事医学科学院吉林大学河北科技师范学院更多>>
发文基金:国家科技重大专项国家农业科技成果转化资金项目更多>>
相关领域:农业科学更多>>

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Construction of DNA Vaccine for FMDV P1 Gene and Immunization Experiment
2013年
[Objective] This study aimed to construct DNA vaccine of foot-and-mouth disease (FMD).[Method] Plasmid carriers plESZP1 and pUTK3CP1 with PRV were constructed for FMDV P1 gene expression.Mice were immunized,and their antibody level was detected.The two eukaryotic expression plasmids constructed were transfected into Vero cells.PCR,IFA and Westem-blot were carried out to detect the transcription and expression of the objective gene.Balb/C mice were intramuscularly inoculated with the DNA plasmid which expressed the target gene correctly,and the antibody level in mice was detected by the means of ELISA and serum neutralization (SN).[Result] DNA plasmid carrying P1 gene which encodes FMDV capsid protein caused specific body fluid immunoreaction in mice,and the antibody level of anti-FMDV had no difference in the mice induced by the two recombinant plasmids.[Conclusion] This study lays a foundation for evaluating the genetically modified vaccine by immunizing animals with recombinant PRV containing the FMDV P1 gene and recombinant virus.
史秋梅高桂生张艳英高光平张东林
关键词:FMDV
高效抗猪瘟病毒shRNA的筛选被引量:1
2014年
为降低RNAi技术的毒副作用,筛选高效抑制猪瘟病毒复制的shRNA序列,根据有关报道构建了荧光报告系统shRNA筛选平台。该平台以9种抑制效率分别为强、中、弱的shRNA筛选载体作为参照,结合荧光报告系统进一步量化shRNA的抑制效率。根据2种已知体外有效抑制猪瘟病毒复制的siRNA-N2、siRNA-NS5B,分别设计得到shRNA-N2、shRNA-NS5B表达载体,继而采用高效、高灵敏度的shRNA筛选平台对这2种shRNA进行了检测,结果显示单拷贝情况下,shRNA-N2抑制靶序列效率较强,shRNA-NS5B抑制效率弱。本试验通过新策略大大提高了shRNA干扰效率检测的灵敏度,为采用shRNA转基因克隆动物进行抗病育种的研究提供了保障。
戴祯王铁东王侃侃欧阳红生涂长春逄大欣
关键词:猪瘟病毒
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