Denatured supercoiled DNA--structural and biological activity.
Supercoiled DNA on treatment with NaOH followed by neutralization produces a condensed structure (form Id). This structure does not split into topoisomers when run on long gel in presence of intercalating agents and the migration of this form does not change appreciably in presence or absence of ethidium bromide. Relaxation of form Id by topoisomerase I from pea chloroplast is facilitated more than form I. Single-stranded binding (SSB) protein binds more to form Id as evidenced from gel retardation study. Hydroxyl radical nicking is facilitated in this form. Compared to form I, this form produces half the number of transformants, but adsorption and penetration remain almost same in both the forms. Post-transformational growth using 32P labelled form I and form Id showed greater amount of degradation in form Id.
Santra
C R
CR
Dept. of Biophysics, Mol. Biol & Genetics, University College of Science, Calcutta, India.
Mukherjee
S K
SK
Thakur
A R
AR
D004278:DNA, Superhelical / Q000737:chemistry* / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D009691:Nucleic Acid Denaturation; D010957:Plasmids / Q000737:chemistry*; D014170:Transformation, Genetic
D004278:DNA, Superhelical