山东农业科学2017，Vol.49Issue(11)：134-137,4.DOI:10.14083/j.issn.1001-4942.2017.11.027
氯化铯密度梯度离心法提纯枣疯病植原体DNA
Separation and Purification of Phytoplasma Genomic DNA of Jujube Witches' Broom by Cesium Chloride Density Gradient Centrifugation
摘要
Abstract
The total DNA was extracted from infected jujube leaves using CTAB method and then the genomic DNA of jujube witches' broom(JWB) phytoplasma was separated and purified from the total DNA by cesium chloride-bisbezimide density gradient centrifugation method.The specific primers aimed to 26S rDNA of jujube and 16S rDNA of JWB phytoplasma were designed to conducted PCR amplification with infected jujube total DNA before and after purification as template respectively.Our findings suggested that when the concentration of template DNA was 1 ng/μL,the electrophoretic band brightness of 26S rDNA of purified phytoplasma DNA was lower than that of total DNA,but the electrophoretic band brightness of JWB was brighter.Moreover,the results of real-time PCR showed that when the concentration of template DNA was 1 ng/μL,the amount of 26S rDNA in the purified phytoplasma DNA relative to that in total DNA was 0.0638,while the amount of JWB relative to that in total DNA was 0.5035.It suggested that the phytoplasma DNA could be effectively isolated and purified from infected jujube total DNA by the method of cesium chloride-bisbezimide density gradient centrifugation.关键词
枣疯病/植原体/CTAB法/氯化铯-双苯酰亚胺密度梯度离心Key words
Jujube witches' broom/Phytoplasma/CTAB method/Cesium chloride-bisbezimide density gradient centrifugation分类
农业科技引用本文复制引用
刘玲雪,肖琦,杨淑珂,路兴波,高瑞..氯化铯密度梯度离心法提纯枣疯病植原体DNA[J].山东农业科学,2017,49(11):134-137,4.基金项目
国家自然科学基金青年基金项目(31401718) （31401718）
山东省优秀中青年科学家科研奖励基金项目(BS2013NY012) （BS2013NY012）
山东省农业科学院青年英才培养计划项目 （）
