Supercondensed structure of plasmid pBR322 DNA in an Escherichia coli DNA topoisomerase II mutant.
An unusual structural component, supercondensed pBR322 DNA, has been found in plasmid pBR322 DNA samples isolated from a DNA topoisomerase II mutant of Escherichia coli, SD108 (topA+, gyrB225). The supercondensed pBR322 DNA moved faster than supercoiled pBR322 DNA as a homogeneous band in agrose gels when the DNA samples were analysed by electrophoresis. The mobility of the supercondensed DNA was not substantially affected by chloroquine intercalation. The supercondensed pBR322 DNA migrated as a high density "third DNA band" when the samples were subjected to caesium chloride/ethidium bromide gradient equilibrium centrifugation. The unusual pBR322 DNA visualized by electron microscopy was a globoid-shaped particle. These observations suggest that the pBR322 plasmid can assume a tertiary structure other than a supercoiled or relaxed structure. DNA topoisomerases may be involved in the supercondensation of plasmid DNA and chromosomal DNA.
Huang
X T
XT
Department of Biology, Nankai University, Tianjin, Peoples Republic of China.
Chen
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D004250:DNA Topoisomerases, Type II / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification / Q000648:ultrastructure; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D008854:Microscopy, Electron; D009154:Mutation; D010957:Plasmids
D004269:DNA, Bacterial; D004250:DNA Topoisomerases, Type II