摘要
以山茶属(Camellia)植物为实验材料,通过在PCR反应液中加入二甲基亚砜(DMSO)来改进ITS区的扩增效果,并对获得的序列进行了相关的比较分析以及系统发育分析。分析结果表明:(1)PCR反应液中添加4%的DMSO能明显提高ITS的扩增效率,并能有效防止ITS假基因的扩增;(2)与功能拷贝相比,ITS假基因序列的ITS1和ITS2区存在多处碱基缺失,GC含量明显偏低,5.8S区二级结构稳定性降低;(3)应用ITS假基因进行种水平的系统发育分析会对其真实的系统关系造成影响,提示进行系统进化研究时应警惕假基因的干扰。
In this study, PCR amplification of nrDNA ITS of Camellia was improved by the addition of DMSO in PCR protocol, the sequence characteristic of ITS were analyzed. The results showed that： （ 1 ） The PCR amplification efficiency of ITS was improved by the addition of 4% DMSO, and the ITS pseudogenes were prevented from being amplified; （2）ITS pseudogenes exhibited a significant deletion in ITS1 and ITS2 region, and displayed lower GC content and unstable secondary structure; （3）ITS pseudogenes could not provided the real phylogenetic relationships, it showed the necessity of great care when used the ITS sequence in evolutionary studies in genus Camellia.
出处
《植物研究》
CAS
CSCD
北大核心
2014年第3期403-408,共6页
Bulletin of Botanical Research
基金
国家质检总局科技计划项目
云南出口花卉物种信息平台建立及分子生物学检测技术研究(2010IK273)
作者简介
尤欢（1988-），女，硕士研究生，主要从事植物学的研究。
通信作者：E-mail：dingdym2008@yahoo．com．cn
