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Retention of helical structure of pBR322 covalently closed circular duplex DNA at high alkaline pH
C R SANTRA, A R THAKUR
Published in BUTTERWORTH-HEINEMANN LTD
1984
Volume: 6
   
Issue: 3
Pages: 152 - 154
Abstract
Denaturation of covalently closed circular duplex replicative form (RF) I at high pH yields a form with high sedimentation coefficient even after neutralization. This form allowed less ethidium bromide to be intercalated but yielded a circular dichroic spectrum which had reduced magnitude of both positive circular dichroism at 273 nm and negative circular dichroism at 245 nm. The circular dichroic spectrum of this form is similar to that of RFV DNA. Gel electrophoretic analysis of this DNA revealed that, although part is retained in the groove, another part appeared as a faster-moving band, which we designated as RF Id. This faster-moving form is cleaved by the restriction endonuclease BamHI at a single site giving a single RF III, comigrating with the RF III obtained from RF I by BamHI cleavage. This signifies that the two strands of RF I did not slide over one another during the formation of RF Id as suggested previously. © 1984.
About the journal
JournalInternational Journal of Biological Macromolecules
PublisherBUTTERWORTH-HEINEMANN LTD
ISSN0141-8130