一种适于PCR检测病毒诱导基因沉默番茄植株的微量DNA提取方法

【研究目的】为了寻找一种适用于病毒诱导的基因沉默番茄叶片PCR检测的快速、简单、成本低的DNA提取方法。【方法】笔者根据PCR反应的要求,用改良的CTAB法,实现了微量番茄叶片基因组DNA的快速提取。【结果】提取基因组DNA所用的组织量虽少,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1.7A,片段大小分别为500bp、1300bp。测序结果证明是相应基因的部分片段。【结论】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。

A Method of Mini-preparation DNA for Gene Silencing Tomato Suited to PCR Detection

OBJECTIVE]In order to seek one kind of DNA extraction method to be suitable for PCR examination of the viral induction gene silence tomato leaf, which is fast, simple and cheap;METHOD]The author according to the requirement of PCR, with the improvement CTAB method, has realized the mini-preparation DNA of tomato leaf; RESULTS]Although the organization quantity which used to extract the genome DNA is few, obtained DNA has degenerated after the electrophoresis, but sufficiently uses in the PCR examination, takes it as the template to expand the DNAm15 and 1.TA of the Su gene silencing plant induced by Tomato yellow leaf curl China virus, the fragment size separately as 500bp, 1300bp. The result of sequence determined is the corresponding gene partial fragments; CONCLUSION]This method material does not need to use the liquid nitrogen, operating independently, and in the gene silence tomato adult plant can stablely and accurately formalizated the PCR examination.