改良一步法提取植物RNA·DNA和蛋白质的研究

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

[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 mays L.)、大豆(Glycine max L.)、苜蓿(Medicago sativa L.)和黄瓜(Cucumis sativus L.)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蛋白质进行鉴定。[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。

[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.