| 舞毒蛾核型多角体病毒PCR检测技术的建立 |
| |
| 引用本文： | 陶婧,韩崇选,张永安,王玉珠,曲良建,王青华.舞毒蛾核型多角体病毒PCR检测技术的建立[J].林业科学研究,2012,25(5):616-619. |
| |
| 作者姓名： | 陶婧 韩崇选 张永安 王玉珠 曲良建 王青华 |
| |
| 作者单位： | 1. 西北农林科技大学林学院,陕西杨凌,712100
2. 中国林业科学研究院森林生态环境与保护研究所,国家林业局森林保护学重点实验室,北京100091 |
| |
| 基金项目： | 林业公益性行业科研专项(200704012,200904029) |
| |
| 摘 要： | 根据舞毒蛾核型多角体病毒(LdMNPV)的蜕皮甾体尿苷二磷酸葡萄糖基转移酶基因(egt)设计一对特异性引物.成功建立了LdMNPV的PCR检测技术体系,其对LdMNPV基因组的灵敏度可达到1fg·mL-1.应用该技术体系从舞毒蛾带毒幼虫、卵、蛹的基因组中扩增出目的片段,证明了这一技术的可行性.对不同浓度的多角体悬液的扩增结果显示:其检测最低量为5 OBs·mL-1.形态学研究表明:从内蒙舞毒蛾体内分离到的LdMNPV中,少数表面有凹陷的孔洞,病毒粒子从这些孔洞中游离出来,这也阐明了此技术能够从多角体悬液中扩增出目的片段的原因.在将来LdMNPV的检测中,可用虫体进行研磨过滤离心得到的组织液作为模板进行扩增.
|
| 关 键 词： | 舞毒蛾核型多角体病毒 PCR 微量检测 egt基因 |
| 收稿时间： | 2012/1/12 0:00:00 |
Establishment of PCR Assay for Detecting of Lymantria dispar Multiple-embedded Nucleopolyhedrovirus (LdMNPV) |
| |
| TAO Jing,HAN Chong-xuan,ZHANG Yong-an,WANG Yu-zhu,QU Liang-jian and WANG Qing-hua.Establishment of PCR Assay for Detecting of Lymantria dispar Multiple-embedded Nucleopolyhedrovirus (LdMNPV)[J].Forest Research,2012,25(5):616-619. |
| |
| Authors: | TAO Jing HAN Chong-xuan ZHANG Yong-an WANG Yu-zhu QU Liang-jian and WANG Qing-hua |
| |
| Institution: | College of Forestry, Northwest A & F University, Yangling 712100, Shaanxi, China;College of Forestry, Northwest A & F University, Yangling 712100, Shaanxi, China;Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry; Key Laboratory of Forest Protection, State Forestry Administration, Beijing 100091, China;Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry; Key Laboratory of Forest Protection, State Forestry Administration, Beijing 100091, China;Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry; Key Laboratory of Forest Protection, State Forestry Administration, Beijing 100091, China;Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry; Key Laboratory of Forest Protection, State Forestry Administration, Beijing 100091, China |
| |
| Abstract: | A technical architecture used for the microdetermination of Lymantria dispar multiple-embedded nucleopolyhedrovirus was set up. Based on egt gene of LdMNPV, a pair of primers was designed with the sensitivity of 1fg·mL-1 DNA. By using this pair of primers, the partial sequences of egt gene were amplified from the eggs, larvae and pupae of the infected L. dispar, which verified the feasibility of this method. The partial sequences of egt gene were also amplified from the polyhedron suspension and the minimum amount of detection could be as low as 5 OBs·mL-1 of polyhedron. Morphology research indicated that the surface of a few of polyhedrons was rough pitted and released virions. It could be a reason why the target DNA fragment could be amplified from the virus suspension. So in the detection of LdMNPV, suspension after trituration, filtration and centrifugalism can be used as template. |
| |
| Keywords: | Lymantria dispar multiple-embedded nucleopolyhedrovirus (LdMNPV) PCR microdetermination egt gene |
| 本文献已被 万方数据 等数据库收录！ |
| 点击此处可从《林业科学研究》浏览原始摘要信息 |
| 点击此处可从《林业科学研究》下载免费的PDF全文 |
|
