摘要
以成熟中苎1号叶片为材料,采用Percoll密度梯度离心法,分离获得完整的叶绿体,并用植物基因组DNA提取试剂盒提取叶绿体DNA,经核酸和蛋白分析仪、琼脂糖电泳、核基因组及叶绿体基因组特异引物PCR检测,结果表明:所提取的苎麻叶绿体DNA纯度高,不含核基因组DNA,并适用于基因扩增等分子操作,为深入研究苎麻叶绿体基因组奠定基础。同时,本方法操作简便、快速,适用性广,为其它物种叶绿体DNA的提取提供借鉴。
Undamaged chloroplast was isolated by using Percoll differential and density -gradient centrifugation from leaf tissues of ramie （ Boehmeria nivea （ L.） Gaud.） , Zhongzhu No.1.And then, Plant Genomic DNA Kit was used to extract chloroplast DNA.The detection results of nucleic acid pro-tein analyzer , agrose gel electrophoresis and PCR amplification based on chloroplast and nuclear genome specific primers showed that ramie cpDNA with high purity and quality , and no nuclear genome DNA might provide a foundation for further study of chloroplast genome in ramie.The DNA was suitable for be-ing used as template in molecular experiment researches such as PCR reaction , gene cloning and so on.The method is rapid , simple and also suitable for isolating cpDNA from many other plants.
出处
《中国麻业科学》
2013年第5期239-243,252,共6页
Plant Fiber Sciences in China
基金
中国农业科学院基本科研业务项目(1610032012030)
关键词
苎麻
叶绿体DNA
密度梯度
提取
Boehmeria nivea （ L.） Gaud.
cpDNA
density-gradient centrifugation
isolation
作者简介
郑建树（1983-），男，博士研究生，主要从事苎麻遗传育种研究。E—mail：zhengjs4142@126．com
通讯作者：熊和平（1955-），男，研究员，主要从事苎麻遗传育种研究。E-mail：ramiexhp@189．cn
