线粒体DNA的异质性与检测方法

线粒体DNA的异质性在生物学、医学、法医、生物技术等领域有重要的应用。本文从自然发生(包括体细胞突变、父系渗入和杂交)和人工产生(包括转基因、细胞融合、核移植和转线粒体)两大方面系统地介绍了线粒体DNA异质性的产生机制及其遗传。并介绍了线粒体DNA异质性的检测方法,已知突变位点mtDNA异质性的检测方法有原位PCR技术、PCR-RFLP和实时荧光定量PCR技术,未知突变位点mtDNA异质性的检测方法有长PCR技术、时相温度梯度凝胶电泳(TTGE)和变性高效液相色谱(DHPLC)等。最后对克隆生物中线粒体异质性检测的应用实例作了介绍。

mtDNA Heteroplasmy and its Detection Methods

The mtDNA heteroplasmy that occurs naturally(somatic mutation,paternal introgression,and hybridization) and mtDNA heteroplasmy that is induced artificially(transgenesis,cell fusion,nuclear transfer,and mitochondrial transfer) as well as the related mechanisms are reviewed.The methods for detecting mtDNA heteroplasmy are described in two categories.mtDNA heteroplasmy with known mutation sites is detected by using in situ PCR,PCR-RFLP,and real-time fluorescent quantitative PCR,while mtDNA heteroplasmy with unknown mutation sites is detected by using long PCR,Temporal Temperature Gradient Gel Electrophoresis(TTGE),and DHPLC.Finally,some examples of detecting mtDNA heteroplasmy in cloned animals are introduced briefly.