Atomic force microscopic study on topological structures of pBR322 DNA

Sci China C Life Sci. 1996 Feb;39(1):1-7.

Abstract

Plasmid pBR322 DNA (0.5 mg/mL) isolated from Escherichia coli HB101 was suspended in Tris-HCI-EDTA (1 mol/L-0.1 mol/L, pH 8.5); then a drop of the above solution was deposited on freshly cleaved mica substrate. After adsorption for about 1 mm, the sample was stained with phosphotungstic acid. The residual solution was removed with a piece of filter paper. Afterwards the sample was imaged with a home-made atomic force microscope (AFM) in air. The AFM images of pBR322 DNA with a molecular resolution have been obtained. These images show that pBR322 DNA exists in several different topological structures: (i) relaxed circular DNA with a different diameter; (ii) supercondensed DNA with different particle sizes; (iii) dimeric catenane connected by one relaxed circular molecule and another close-compacted molecule which might be either supercoiled or intramolecular knotted form; (iv) oligomeric catenane with multiple irregular molecules in which DNA is interlocked into a complex oligomer; (v) possibly-existing intermediates formed from monomer to oligomer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Bacterial / isolation & purification
  • DNA, Bacterial / ultrastructure*
  • Escherichia coli / chemistry
  • Escherichia coli / genetics
  • Microscopy, Atomic Force
  • Plasmids / isolation & purification
  • Plasmids / ultrastructure*

Substances

  • DNA, Bacterial