Melov S, Lithgow G J, Fischer D R, Tedesco P M, Johnson T E
Institute for Behavioral Genetics, University of Colorado, Boulder 80309, USA.
Nucleic Acids Res. 1995 Apr 25;23(8):1419-25. doi: 10.1093/nar/23.8.1419.
We have developed a long-extension-PCR strategy which amplifies approximately half of the mitochondrial genome (6.3 kb) of Caenorhabditis elegans using an individual worm as target. We analyzed three strains over their life span to assess the number of detectable deletions in the mitochondrial genome. Two of these strains are wild-type for life span while the third is mutant in the age-1 gene, approximately doubling its maximum life span. At the mean life span in wild-type strains, there was a significant difference between the frequency of deletions detected in the mitochondrial genome compared with the mean number of deletions in young animals. In addition, deletions in the mitochondrial genome occur at a significantly lower rate in age-1 mutants as compared with wild type. We cloned and identified the breakpoints of two deletions and found that one of the deletions had a direct repeat of 8 bp at the breakpoint. This is the largest single study (over 900 individual animals) characterizing the frequency of deletions in the mitochondrial genome as a function of age yet carried out.
我们开发了一种长延伸聚合酶链反应策略，该策略以单个线虫为靶点，扩增秀丽隐杆线虫线粒体基因组的大约一半（6.3 kb）。我们分析了三个品系在其整个生命周期中的线粒体基因组中可检测到的缺失数量。其中两个品系的寿命为野生型，而第三个品系在age-1基因中发生了突变，其最大寿命大约延长了一倍。在野生型品系的平均寿命时，线粒体基因组中检测到的缺失频率与幼龄动物的平均缺失数之间存在显著差异。此外，与野生型相比，age-1突变体中线粒体基因组的缺失发生率显著降低。我们克隆并鉴定了两个缺失的断点，发现其中一个缺失在断点处有一个8 bp的直接重复序列。这是迄今为止针对线粒体基因组中缺失频率随年龄变化所进行的最大规模的单项研究（超过900只个体动物）。
