通过宿主细菌的低温培养降低克隆的真核串联重复DNA的结构不稳定性
Reduction in the structural instability of cloned eukaryotic tandem-repeat DNA by low-temperature culturing of host bacteria.
作者信息
Thapana Watcharaporn, Sujiwattanarat Penporn, Srikulnath Kornsorn, Hirai Hirohisa, Koga Akihiko
机构信息
Primate Research Institute, Kyoto University,Inuyama City 484-8506,Japan.
Faculty of Science,Kasetsart University,Bangkok 10900,Thailand.
出版信息
Genet Res (Camb). 2014 Oct 27;96:e13. doi: 10.1017/S0016672314000172.
Summary For accurate analyses of eukaryotic tandem-repeat DNA, it is often required to clone a genomic DNA fragment into a bacterial plasmid. It is, however, a serious problem that tandem-repeat DNA is frequently subjected to structural changes during maintenance or amplification in the host bacteria. Here, we show an example of a clear difference in the instability of tandem-repeat DNA between different culturing temperatures. A fragment of monkey centromeric DNA carried by pUC19 was considerably degraded by culturing bacteria at 37 °C, but the damage was reduced at 25 °C. Thus, culturing temperature is a significant factor for avoiding degradation, in addition to the genotype of the host bacteria.
摘要 为了准确分析真核生物串联重复DNA，通常需要将基因组DNA片段克隆到细菌质粒中。然而，一个严重的问题是，串联重复DNA在宿主细菌中维持或扩增期间经常发生结构变化。在这里，我们展示了一个不同培养温度下串联重复DNA不稳定性存在明显差异的例子。携带在pUC19上的猴着丝粒DNA片段在37°C培养细菌时会被大量降解，但在25°C时损伤会减少。因此，除了宿主细菌的基因型外，培养温度也是避免降解的一个重要因素。
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