title: Reduction in the structural instability of cloned eukaryotic tandem-repeat DNA by low-temperature culturing of host bacteria.
abstract: 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.
authors.affiliation: Primate Research Institute, Kyoto University,Inuyama City 484-8506,Japan.
authors.forename: Watcharaporn
authors.initials: W
authors.lastname: Thapana
authors.affiliation: Primate Research Institute, Kyoto University,Inuyama City 484-8506,Japan.
authors.forename: Penporn
authors.initials: P
authors.lastname: Sujiwattanarat
authors.affiliation: Faculty of Science,Kasetsart University,Bangkok 10900,Thailand.
authors.forename: Kornsorn
authors.initials: K
authors.lastname: Srikulnath
authors.affiliation: Primate Research Institute, Kyoto University,Inuyama City 484-8506,Japan.
authors.forename: Hirohisa
authors.initials: H
authors.lastname: Hirai
authors.affiliation: Primate Research Institute, Kyoto University,Inuyama City 484-8506,Japan.
authors.forename: Akihiko
authors.initials: A
authors.lastname: Koga
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics / Q000254:growth & development*; D018929:Cell Culture Techniques / Q000379:methods*; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D018014:Gene Transfer Techniques; D042822:Genomic Instability / Q000235:genetics*; D000882:Haplorhini; D010957:Plasmids / Q000235:genetics*; D020080:Tandem Repeat Sequences / Q000235:genetics*; D013696:Temperature