Reduction in the structural instability of cloned eukaryotic tandem-repeat DNA by low-temperature culturing of host bacteria
Reduction in the structural instability of cloned eukaryotic tandem-repeat DNA by low-temperature culturing of host bacteria
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DOI：
10.1017/s0016672314000172
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发表时间：
2014-01-01
影响因子：
1.5
通讯作者：
Koga, Akihiko
中科院分区：
文献类型：
--
作者：
Thapana, Watcharaporn;Sujiwattanarat, Penporn;Koga, Akihiko
为了准确分析真核生物的串联重复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，我们展示了不同培养温度之间串联重复DNA不稳定性的明显差异。pUC 19携带的猴着丝粒DNA片段在37 ℃培养细菌时被显著降解，但在25 ℃时损伤减少。因此，除了宿主细菌的基因型之外，培养温度是避免降解的重要因素。
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 degrees C, but the damage was reduced at 25 degrees C. Thus, culturing temperature is a significant factor for avoiding degradation, in addition to the genotype of the host bacteria.