摘要
目的：对麻花秦艽和粗茎秦艽叶绿体基因组进行提取研究，为进一步研究秦艽叶绿体的基因组序列特征、遗传多样性以及资源保护研究奠定基础。方法：本文以麻花秦艽、粗茎秦艽为对象，采用改良的蔗糖密度梯度离心方法进行提取，优化纯化方法，分离得到秦艽叶绿体后测定cpDNA浓度，检测其OD值并扩增ITS2序列验证其纯度。结果：获得高质量、高纯度的cpDNA（0．1～0．4μg／10g），扩增ITS2序列验证其无核污染，满足后续测序要求。结论：cpDNA的纯度和质量对叶绿体基因组测序的正确率和可信度具有重要的影响。本文获得了高纯度、高质量的秦艽cpDNA，为秦艽叶绿体基因组全序列的获得提供保障。
This study was aimed to show the extraction research of cpDNA in Gentiana straminea Maxim. and G. crassicaulis Duthie ex Burk., which will increase the knowledge about the chloroplast genome sequence characteris- tics, understand its genetic diversity and improve the efficiency of breeding schemes. G. straminea Maxim. and G. crassicaulis Duthie ex Burk. were taken as study objects. The improved sucrose-gradient centrifugation and purifica- tion methods were used to obtain the cpDNA, measure the concentration, detect the OD value, and amplify the ITS2 sequence to verify the nucleus pollution. The results showed that the high quality and purity cpDNA （0.1 - 0.4 μg/ 10 g） was demonstrated without other pollution after ITS2 sequence amplification, which met to the subsequent effi- cient sequencing of chloroplast genomes. The purity and mass have an important influence on the accuracy and cred- ibility of the cp genome sequencing. It was concluded that this study improved the sucrose-gradient centrifugation and purification with great effect of the purity and mass of cpDNA, and guaranteed the obtaining of complete cpDNA of Gentiana.
出处
《世界科学技术-中医药现代化》
北大核心
2013年第9期1871-1875,共5页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
科学技术部国家高技术研究发展计划(“863计划”)项目(2012AA021602):基于DNA条形码的珍稀药用、野生等资源快速检测技术及产品研发,负责人:陈士林
教育部长江学者和创新团队发展计划(IRT1150):中药资源学,负责人:陈士林
关键词
麻花秦艽
粗茎秦艽
叶绿体
DNA
提取
G. straminea Maxim., G. crassicaulis Duthie ex Burk., cpDNA, extraction
