Modified One Step Isolation of RNA,DNA and Protein from Plant Tissues  

改良一步法提取植物RNA、DNA和蛋白质的研究(英文)

作　　者：肖笑[1] 韩斌[1] 徐晟[1] 李乐[1] 何健[1] 黄明[2] 谢贵水[3] 沈文飚[1] 

机构地区：[1]南京农业大学生命科学学院,江苏南京210095 [2]南京农业大学食品科技学院,江苏南京210095 [3]中国热带农业科学院橡胶研究所,海南儋州571737

出　　处：《Agricultural Science & Technology》2010年第9期58-62,共5页农业科学与技术（英文版）

基　　金：Supported by Opening Program of Physiologial Key Laboratory for Cultivation of Tropical Crops in Hainan Province(Key Opening Bio-logial Laboratory of Rubber of Ministry of Agriculture/Breeding Basein State and Provincial Key Laboratory)(KLOF0907);Major and Special Program for Cultivation of Genetically Modified New Varieties(No.2009ZX08012-014B);National Natural Science Foundation of China(No.30971711);Natural Science Foundation in Jiangsu Province(No.BK2009309)~~

摘　　要：[Objective] The aim of this study was to extract RNA,DNA and protein from plants by using modified one step method.[Method]CTAB was used for the modifying of Biozol-based one step extraction method,which was then used for the isolation of RNA,DNA and protein from the roots of maize（Zea mays L.）,soybean（Glycine max L.）,alfalfa（Medicago sativa L.）and cucumber（Cucumis sativus L.）.And the heme oxygenase-1 gene（HO-1）and calcium-dependent protein kinase-1 gene（CDPK1）,as well as HO-1 protein were identified.[Result]The result of UV spectrophotometer determination and agarose gel electrophoresis analysis showed that the purity of obtained RNA and DNA sample was relatively high;HO-1 and CDPK1 gene exhibited the positive signals checked by RT-PCR;the extracted protein could be used for western blotting analysis;the whole isolation procedure cost 3 h.[Conclusion]This method can be used as a good reference for researcher and will offer a better value with a wide range of applications.[目的]利用改良的一步法提取植物RNA、DNA和蛋白质。[方法]采用CTAB试剂对Biozol抽提法进行改良,用一步法分别对玉米(Zea maysL.)、大豆(Glycine maxL.)、苜蓿(Medicago sativaL.)和黄瓜(Cucumis sativusL.)4种作物根的RNA、DNA和蛋白质进行共提取,并对其HO-1和CDPK1基因及HO-1蛋白进行鉴定。[结果]经紫外分光光度计测定和琼脂糖凝胶电泳分析证明获得的RNA和DNA样品纯度较高;HO-1基因和CDPK1基因的RT-PCR结果呈阳性;所提取的蛋白质也可用于Western blot分析;整个提取过程只需3h。[结论]该方法实用性较强,具有广泛的应用价值。

关 键 词：One step method Biozol RNA DNA PROTEIN 

分 类 号：Q943.2[生物学—植物学]