Use of high throughput sequencing to observe genome dynamics at a single cell level
Use of high throughput sequencing to observe genome dynamics at a single cell level
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DOI：
10.1073/pnas.0906681106
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发表时间：
2009-12-08
影响因子：
11.1
通讯作者：
Ogryzko, V.
中科院分区：
文献类型：
--
作者：
Parkhomchuk, D.;Amstislavskiy, V.;Ogryzko, V.
随着高通量测序技术的发展，以全基因组的方式直接分析突变分布成为可能，使突变率测量从传统的潜在假设中解脱出来。在这里，我们对化学诱变后获得的菌落中的几个大肠杆菌基因组进行了测序，并观察到诱导突变的非随机分布。这些因素包括沿基因组的G到A或C到T转变的长时间延伸，以及基因组内和基因组间突变密度的数量级差异。虽然这些观察结果中的大多数可以用酶过程的已知特征来解释，但其他的可以在单细胞水平上反映分子过程中的随机性。我们的结果表明，对单个诱变细胞后代基因组中留下的分子记录的分析如何允许在基因组范围内观察其祖先的单个基因组中突变的固定和分离以及重组事件。
With the development of high throughput sequencing technology, it becomes possible to directly analyze mutation distribution in a genome-wide fashion, dissociating mutation rate measurements from the traditional underlying assumptions. Here, we sequenced several genomes of Escherichia coli from colonies obtained after chemical mutagenesis and observed a strikingly nonrandom distribution of the induced mutations. These include long stretches of exclusively G to A or C to T transitions along the genome and orders of magnitude intra- and intergenomic differences in mutation density. Whereas most of these observations can be explained by the known features of enzymatic processes, the others could reflect stochasticity in the molecular processes at the single-cell level. Our results demonstrate how analysis of the molecular records left in the genomes of the descendants of an individual mutagenized cell allows for genome-scale observations of fixation and segregation of mutations, as well as recombination events, in the single genome of their progenitor.