|
|||||
|
|
| 靶向VP1和VP2双拷贝shRNA重组表达质粒抑制传染性法氏囊病病毒复制的研究 | |
| 引用本文： | 欧阳伟,王永山,马金荣,张海彬. 靶向VP1和VP2双拷贝shRNA重组表达质粒抑制传染性法氏囊病病毒复制的研究[J]. 中国预防兽医学报, 2011, 33(10). DOI: 10.3969/j.issn.1008-0589.2011.10.01 |
| 作者姓名： | 欧阳伟 王永山 马金荣 张海彬 |
| 作者单位： | 1. 江苏省农业科学院兽医研究所/国家兽用生物制品工程技术研究中心,江苏南京,210014 2. 江苏省农业科学院兽医研究所/国家兽用生物制品工程技术研究中心,江苏南京210014;南京农业大学动物医学院,江苏南京210095 3. 南京农业大学动物医学院,江苏南京,210095 |
| 基金项目： | 江苏省自然科学基金(BK2008352、BK2010471) |
| 摘 要： | 为抑制传染性法氏囊病病毒(IBDV)的复制,本研究构建了靶向IBDV VP1和VP2基因的鸡双向U6启动子(chU6)双拷贝shRNA重组表达质粒pchU6-shRNA12,将其转染鸡胚成纤维细胞(CEF),再接种IBDV,72 h后,未出现细胞病变(CPE),病毒滴度小于101 TCID50/0.1mL；而转染空质粒和未转染两个对照组均出现明显的CPE,病毒滴度均达到108.75 TCID50/0.1 mL.此外,将pchU6-shRNA12与IBDV混合,接种于10日龄SPF鸡胚尿囊腔,96 h后,鸡胚发育正常,病毒滴度小于101 ELD5,0/0.1 mL,实时荧光定量RT-PCR检测VP1和VP2基因,比单纯接种IBDV组分别降低92％和95％；而含有空质粒和不含质粒两个对照组鸡胚则全部死亡,病毒滴度均达到107.00 ELD50/0.1mL.本研究结果表明,chU6启动子在双拷贝shRNA表达质粒pchU6-shRNA12中能高效驱动双拷贝shRNA的转录,生成的siRNA在CEF(in vitro)和鸡胚体内(in vivo)均能有效抑制IBDV的复制. |
| 关 键 词： | 传染性法氏囊病病毒 RNA干扰 小发夹RNA 鸡U6启动子 |
Inhibition of infectious bursal disease virus replication by duplex shRNAs targeting the VP1 and VP2 genes driven by chicken U6 promoter |
|
| OUYANG Wei,WANG Yong-shan,MA Jin-rong,ZHANG Hai-bin. Inhibition of infectious bursal disease virus replication by duplex shRNAs targeting the VP1 and VP2 genes driven by chicken U6 promoter[J]. Chinese Journal of Preventive Veterinary Medicine, 2011, 33(10). DOI: 10.3969/j.issn.1008-0589.2011.10.01 | |
| Authors: | OUYANG Wei WANG Yong-shan MA Jin-rong ZHANG Hai-bin |
| Affiliation: | OUYANG Wei 1,WANG Yong-shan 1,MA Jin-rong 1,2,ZHANG Hai-bin 2 (1.National Center for Engineering Research of Veterinary Bio-products,Institute of Veterinary Medicine,Jiangsu Academy of Agricutural Sciences,Nanjing 210014,China,2.College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China) |
| Abstract: | To inhibit the replication of infectious bursal disease virus(IBDV),two short hairpin RNAs(shRNAs) targeting VP1 and VP2 genes of IBDV were designed and synthesized.The recombinant plasmid pchU6-shRNA12 containing chicken U6 promoters on each side of the shRNAs based on pSilencer2.1-U6 vector was constructed.In the IBDV inhibition test(in vitro) of the chicken embryo fibroblasts(CEF) infected with IBDV at 24 hours post transfecting with pchU6-shRNA12,the virus titer was less than 10 TCID 50 /0.1 mL,in the c... |
| Keywords: | infectious bursal disease virus RNA interference short hairpin RNA chicken U6 promoter |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |