Establishment of a heteroplasmic mouse strain with interspecific mitochondrial DNA haplotypes and improvement of a PCR-RFLP-based measurement system for estimation of mitochondrial DNA heteroplasmy
Establishment of a heteroplasmic mouse strain with interspecific mitochondrial DNA haplotypes and improvement of a PCR-RFLP-based measurement system for estimation of mitochondrial DNA heteroplasmy
复制标题
具有种间线粒体DNA单倍型的异质性小鼠品系的建立以及基于PCR-RFLP的线粒体DNA异质性评估测量系统的改进
DOI：
10.1007/s11248-017-0009-2
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
L. Cao, M. Yamaguchi, H. Yonekawa、C. Taya
中科院分区：
文献类型：
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作者：
H. Shitara;L. Cao, M. Yamaguchi, H. Yonekawa、C. Taya
线粒体DNA分离是线粒体遗传的特征模式之一，其中线粒体DNA的异质状态以可变的比例传递给下一代。为了分析线粒体DNA的分离，我们制备了一个具有种间线粒体DNA单倍型的异质小鼠品系，该品系包含来自C57 BL/6 J和小家鼠品系的两种类型的线粒体DNA。该菌株是通过将供体细胞质显微注射到受精卵中，在C57 BL/6 J核基因组背景下产生的。采用改进的PCR-RFLP半定量分析方法，减少异源双链的影响，测定组织中线粒体DNA异质性比例。创始小鼠的尾部含有约14%的外源小家鼠线粒体DNA分子，并且同一个体的不同组织中检测到的比例不同。在母系遗传模式下，异质性线粒体DNA以不同的比例传递给下一代。这种线粒体异质性小鼠品系和改进的PCR-RFLP测量系统使得能够分析异质性线粒体DNA变体在组织和世代之间的传递。
Mitochondrial DNA segregation is one of the characteristic modes of mitochondrial inheritance in which the heteroplasmic state of mitochondrial DNA is transmitted to the next generation in variable proportions. To analyze mitochondrial DNA segregation, we produced a heteroplasmic mouse strain with interspecific mitochondrial DNA haplotypes, which contains two types of mitochondrial DNA derived from C57BL/6J andMus spretusstrains. The strain was produced on a C57BL/6J nuclear genomic background by microinjection of donor cytoplasm into fertilized eggs. The PCR-RFLP semi-quantitative analysis method, which was improved to reduce the effect of heteroduplex formation, was used to measure the proportion of heteroplasmic mitochondrial DNA in tissues. Founder mice contained up to approximately 14% of exogenousMus spretusmitochondrial DNA molecules in their tails, and the detected proportions differed among tissues of the same individual. Heteroplasmic mitochondrial DNA is transmitted to the next generation in varying proportions under the maternal inheritance mode. This mitochondrial heteroplasmic mouse strain and the improved PCR-RFLP measurement system enable analysis of the transmission of heteroplasmic mitochondrial DNA variants between tissues and generations.