| 一种快速提取烟草病毒的方法 |
| |
| 引用本文： | 谷瑞华,王进忠,文思远,王升启,张爱环,孙淑玲,赵世恒.一种快速提取烟草病毒的方法[J].北京农学院学报,2006,21(4):68-70. |
| |
| 作者姓名： | 谷瑞华 王进忠 文思远 王升启 张爱环 孙淑玲 赵世恒 |
| |
| 作者单位： | 北京农学院,植物科学技术系,北京,102206;军事医学科学院放射医学研究所,北京,100850 |
| |
| 基金项目： | 北京市教委会资助项目(KM200610020007)，北京市自然基金资助项目(5043026)，北京市市管高校人才强教计划资助项目 |
| |
| 摘 要： | 为寻找高效快速提取植物总RNA的检测方法,以白肋烟叶片为材料,用Trizol试剂盒提取总RNA。依据黄瓜花叶病毒和马铃薯X病毒的外壳蛋白保守序列设计特异性引物,进行RT-PCR检测,从感病叶片中分别扩增出326bp、392bp的目的片断,而对照叶片中无此扩增带。将PCR产物连接pGEM-T载体上,转化大肠杆菌DH5α,得到了含有目的片断的重组子。序列分析表明,与Gene Bank中报道的这两种病毒核苷酸序列进行比较,同源性均达到96%以上。从而证明Trizol试剂盒可以在1 h内提取到纯度高、含量高、完整性好的总RNA,快速、简便、可靠。
|
| 关 键 词： | 黄瓜花叶病毒 马铃薯X病毒 RT-PCR Trizol试剂盒 |
| 文章编号： | 1002-3186(2006)04-0068-03 |
| 收稿时间： | 2006-08-30 |
| 修稿时间： | 2006-09-30 |
A Rapid Method for the Extraction Viruses in Tobacco |
| |
| GU Rui-hua,WANG Jin-zhong,WEN Si-yuan,WANG Sheng-qi,ZHANG Ai-huan,SUN Shu-ling,ZHAO Shi-heng.A Rapid Method for the Extraction Viruses in Tobacco[J].Journal of Beijing Agricultural College,2006,21(4):68-70. |
| |
| Authors: | GU Rui-hua WANG Jin-zhong WEN Si-yuan WANG Sheng-qi ZHANG Ai-huan SUN Shu-ling ZHAO Shi-heng |
| |
| Institution: | 1. Department of Plant Science and Technology, BeiJing Agricultural College, Beijing 102206 ,China;2. Radiation Medicine,Academy of Military Medical Science,Beijing 100850,China |
| |
| Abstract: | Total RNA from the leaves of N.tabacum cv.White burly was extracted by Trizol Reagent.Using the method,it was obtained the total RNA with the high purity,high quality and integrality in 1 h.Special primers of cucumber mosaic virus and potato X virus were designed respectively.The expected sizes of 326bp and 392bp were amplified by RT-PCR separately from the infected sample,but not from the control sample.The PCR products were ligated to pGEM-T Vector,sequencing validated their identities to the consensus sequence in the GenBank.The homology both achieve above 96%.That method is suitable for detecting RNA viruses by RT-PCR rapidly,simply,and reliably. |
| |
| Keywords: | cucumber mosaic virus potato X virus RT-PCR Trizol Reagent box |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
