农业科学与技术(英文版)
農業科學與技術(英文版)
농업과학여기술(영문판)
AGRICULTURAL SCIENCE & TECHNOLOGY
2010年
9期
58-62
,共5页
肖笑%韩斌%徐晟%李乐%何健%黄明%谢贵水%沈文飚
肖笑%韓斌%徐晟%李樂%何健%黃明%謝貴水%瀋文飚
초소%한빈%서성%리악%하건%황명%사귀수%침문표
一步法%Biozol%RNA%DNA%蛋白质
一步法%Biozol%RNA%DNA%蛋白質
일보법%Biozol%RNA%DNA%단백질
One step method%Biozol%RNA%DNA%Protein
[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 3 h.[结论]该方法实用性较强,具有广泛的应用价值.
[目的]利用改良的一步法提取植物RNA、DNA和蛋白質.[方法]採用CTAB試劑對Biozol抽提法進行改良,用一步法分彆對玉米(Zea mays L.)、大豆(Glycine max L.)、苜蓿(Medicago sativa L.)和黃瓜(Cucumis sativus L.)4種作物根的RNA、DNA和蛋白質進行共提取,併對其HO-1和CDPK1基因及HO-1蛋白進行鑒定.[結果]經紫外分光光度計測定和瓊脂糖凝膠電泳分析證明穫得的RNA和DNA樣品純度較高;HO-1基因和CDPK1基因的RT-PCR結果呈暘性;所提取的蛋白質也可用于Western blot分析;整箇提取過程隻需 3 h.[結論]該方法實用性較彊,具有廣汎的應用價值.
[목적]이용개량적일보법제취식물RNA、DNA화단백질.[방법]채용CTAB시제대Biozol추제법진행개량,용일보법분별대옥미(Zea mays L.)、대두(Glycine max L.)、목숙(Medicago sativa L.)화황과(Cucumis sativus L.)4충작물근적RNA、DNA화단백질진행공제취,병대기HO-1화CDPK1기인급HO-1단백진행감정.[결과]경자외분광광도계측정화경지당응효전영분석증명획득적RNA화DNA양품순도교고;HO-1기인화CDPK1기인적RT-PCR결과정양성;소제취적단백질야가용우Western blot분석;정개제취과정지수 3 h.[결론]해방법실용성교강,구유엄범적응용개치.
[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.