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植物研究 ›› 2008, Vol. 28 ›› Issue (6): 726-729.doi: 10.7525/j.issn.1673-5102.2008.06.017

• 论文 • 上一篇    下一篇

有效提取病毒诱导基因沉默植株中DNA的方法

何秀霞;于源华;果洪宇;张淑华   

  1. (长春理工大学生命科学技术学院,长春 130022)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-11-20 发布日期:2008-11-20
  • 基金资助:
     

A Effective Preparation Method for DNA from Gene Silencing Plant Induced by Virus

HE Xiu-Xia;YU Yuan-Hua;GUO Hong-Yu;ZHANG Shu-Hua   

  1. (College of Life Science,Changchun University of Science and Technology,Changchun 130022)
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-11-20 Published:2008-11-20
  • Supported by:
     

摘要: 根据PCR反应的要求,用改良的CTAB法,以番茄植株为材料,实现了微量番茄叶片基因组DNA的快速提取。提取基因组DNA所用的组织量少,所得的DNA经过电泳检测虽有降解,但足以用于PCR检测,以其作模板扩增中国番茄黄化曲叶病毒诱导的硫黄素酶(Su)基因沉默植株中病毒组分中的DNAmβ和1.7 A,片段大小分别为1 300、500 bp。测序结果证明是相应基因的部分片段。该方法的材料不需要使用液氮,可以单人大批量提取,并在基因沉默的番茄植株中能稳定而准确的规模化PCR检测。

关键词: PCR, DNA微量提取方法, 番茄

Abstract: According to the requirement of PCR, with the improved CTAB method, taking tomato as material, the minipreparation DNA of tomato leaf was realized; Although the organization quantity which used to extract the genome DNA is few, and the genome DNA has degenerated after the electrophoresis, but was sufficient for uses in the PCR examination. taking it as the template to amplify the DNAmβ and 1.7A of the Su gene silencing plant induced by Tomato yellow leaf curl China virus, the fragment size was 1 300 bp and 500 bp,respectively. The result of sequence determined is the corresponding gene partial fragments. This method does not need to use liquid nitrogen, can operate independently, and can stablely and accurately formalizated the PCR examination in the gene silence tomato plant.

Key words: PCR, method of mini-preparation DNA, tomato

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