Reduction in the structural instability of cloned eukaryotic tandem-repeat DNA by low-temperature culturing of host bacteria

Thapana W, Sujiwattanarat P, Srikulnath K, Hirai H and *Koga A

タパナ ワチャラポン・スジワッタナラ ペンポン、コンソン シクルナ、平井啓久、*古賀章彦

真核生物の反復配列を正確に分析するためには、ゲノムＤＮＡの断片のクローン化が必要になることが多い。しかし、クローンをホストのバクテリアで維持または増幅している崩壊することがよくあり、正確な分析の妨げとなっている。バクテリアを低温で培養することで崩壊が軽減できることを、実例とともに示す。

Genetics Research 96: e13 (2014)

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 37oC, but the damage was reduced at 25oC. Thus, culturing temperature is a significant factor for avoiding degradation, in addition to the genotype of the host bacteria

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