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Construction of Short Hairpin RNA Vector with σNS & σC Genes of Avian Reovirus and Determination of Interference Effect
  • ISSN号:0366-6964
  • 期刊名称:《畜牧兽医学报》
  • 时间:0
  • 分类:Q782[生物学—分子生物学] Q522[生物学—生物化学]
  • 作者机构:[1]Guangxi Veterinary Research Institute ,Guangxi Key Laboratory of Animal Vaccines and New Technology, Nanning 530001, China.
  • 相关基金:This work was supported by National Natural Science Foundation (31160512), Funds for Special Expert of Guangxi Province (2011 B020 ) and Guangxi Science and Techology Research Projects (0991222).
中文摘要:

[Objective] The aim was to explore novel method for treatment of Avian Reovirus. [Method] According to the design principle of siRNA target sequences, siRNA templates were designed and synthesized and then cloned into the shRNA expression vector, namely, pSilencer-CMV 4.1 neo. Short hairpin RNA vector C1, C2, C3, which contain σC gene, and shRNA vector NS1, NS2, NS3, which contain σNS gene, were constructed separately. The constructed shRNA vectors and negative control were co-transfected into DF-1 cells with the eukaryotic expression vector pEGFP-σC and pEGFP-σNS, respectively. [Result] Observation through fluorescence microscope indicated that the constructed 6 shRNA could inhibit the expression of fusion protein to different degrees. In addition, results of Real-time PCR suggested that C3 and NS1 have the best interference effect to the viral duplication in vitro. [Conclusion] Construction and selection of specific shRNA expression vectors inhibiting Avian Reovirus are significant for researching effects of σC and σNS proteins in infection and duplication of ARV, providing new idea for ARV antiviral therapy.

英文摘要:

[ Objective] The aim was to explore novel method for treatment of Avian Reovirus. [ Method] According to the design principle of siRNA target sequences, siRNA templates were designed and synthesized and then cloned into the shRNA expression vector, namely, pSilencer-CMV 4.1 neo. Short hairpin RNA vector C1, C2, C3, which contain σC gene, and shRNA vector NS1, NS2, NS3, which contain aNS gene, were constructed separately. The constructed shRNA vectors and negative control were co-transfected into DF-1 cells with the eukaryotic expression vector pEG- FP-σC and pEGFP-σNS, respectively. [ Result] Observation through fluorescence microscope indicated that the constructed 6 shRNA could inhibit the expression of fusion protein to different degrees. In addition, results of Real-time PCR suggested that C3 and NS1 have the best interference effect to the viral duplication in vitro. [ Conclusionl Construction and selection of specific shRNA expression vectors inhibiting Avian Reovirus are significant for researching effects of σC and oNS proteins in infection and duplication of ARV, providing new idea for ARV antiviral therapy.

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期刊信息
  • 《畜牧兽医学报》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学技术协会
  • 主办单位:中国畜牧兽医学会
  • 主编:文杰
  • 地址:北京海淀区圆明园西路2号中国农科院畜牧所
  • 邮编:100193
  • 邮箱:xmsyxb@263.net
  • 电话:010-62815987 62816996
  • 国际标准刊号:ISSN:0366-6964
  • 国内统一刊号:ISSN:11-1985/S
  • 邮发代号:82-453
  • 获奖情况:
  • 1992年北京优秀期刊奖,1998年、2000年在全国畜牧兽医优秀期刊评比中获一等奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,波兰哥白尼索引,美国剑桥科学文摘,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),瑞典开放获取期刊指南,中国北大核心期刊(2000版)
  • 被引量:21857