| 靶向猪瘟病毒NS3基因shRNA干扰载体的构建与鉴定 |
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| 引用本文： | 冶贵生,张彦明,徐 浩.靶向猪瘟病毒NS3基因shRNA干扰载体的构建与鉴定[J].西北农林科技大学学报(社会科学版),2007,35(3):7-10. |
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| 作者姓名： | 冶贵生 张彦明 徐 浩 |
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| 作者单位： | 西北农林科技大学,动物科技学院,陕西,杨凌,712100 |
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| 基金项目： | 国家自然科学基金;西北农林科技大学校科研和教改项目 |
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| 摘 要： | 利用RNA干扰在线生物学软件分析猪瘟病毒NS3基因,设计针对NS3基因不同位置的小发卡RNA(short hairpin RNA,shRNA)干扰序列(其结构特征为正链(19nt)-环(9nt)-负链(19nt))。化学合成这些序列,并退火连接为双链干扰片段,将双链干扰片段定向克隆到干扰载体中,构建重组载体pGene-NS3-1,pGene-NS3-2和pGene-NS3-3,转化DH5α大肠杆菌感受态细胞,对转化产物进行kan+筛选,碱裂解法提取重组质粒,将酶切鉴定为阳性的质粒测序。结果显示,双链干扰片段克隆方向正确,序列中未出现核苷酸的插入,缺失等突变现象。表明靶向猪瘟病毒NS3基因shRNA干扰载体构建成功。
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| 关 键 词： | 猪瘟病毒 NS3基因 RNA干扰 小发卡RNA |
| 文章编号： | 1671-9387（2007）03-0007-04 |
| 收稿时间： | 2006/2/17 0:00:00 |
| 修稿时间： | 2006-02-17 |
Construction and identification of recombinant shRNA interference plasmids targeting to NS3 gene of classic swine fever virus |
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| YE Gui-sheng,ZHANG Yan-ming,XU Hao,GUO Kang-kang.Construction and identification of recombinant shRNA interference plasmids targeting to NS3 gene of classic swine fever virus[J].Journal of Northwest Sci-Tech Univ of Agr and,2007,35(3):7-10. |
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| Authors: | YE Gui-sheng ZHANG Yan-ming XU Hao GUO Kang-kang |
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| Institution: | (Colleage of Animal Science and Veterinary Medicine,Northwest A & F University,Yangling,Shaanxi 712100,China) |
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| Abstract: | 19nt interferencing sequences for selecting different positions of NS3 gene were designed for Sence-Loop-Antisence by using biology software after analysising NS3 gene of classic swine fever virus.Then,these sequences of chemical synthesis were annealed for double strands interference segment,which was made directional cloning to interferecing plasmid to construct recombinat plasmids pGene-NS31,pGene-NS3-2 and p-Gene-NS3.The results indicated that segments were cloned rightly and there wasn't nucleotide insertion and nucleotide deletion,and recombinant shRNA interference plasmids targeting to NS3 gene of CSFV was successfully constructed,which was basic for researching NS gene. |
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| Keywords: | classical swine fever virus NS3 gene RNA interference small hairpin RNA |
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