Correction: Determination of the Number of Superhelical Turns in Simian Virus 40 DNA by Gel Electrophoresis

1976 ◽  
Vol 73 (7) ◽  
pp. 2527-2527
Keyword(s):  
Gel Electrophoresis ◽  
Simian Virus 40 ◽  
Simian Virus

Two-dimensional gel electrophoretic method for mapping DNA replicons

1988 ◽  
Vol 8 (4) ◽  
pp. 1408-1413 ◽  
Author(s):  
K A Nawotka ◽  
J A Huberman
Keyword(s):  
Gel Electrophoresis ◽  
Replication Origin ◽  
Replication Fork ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Two Dimensional ◽  
Agarose Gel Electrophoresis ◽  
Electrophoretic Method ◽  
Short Probe

We describe in detail a method which allows determination of the directions of replication fork movement through segments of DNA for which cloned probes are available. The method uses two-dimensional neutral-alkaline agarose gel electrophoresis followed by hybridization with short probe sequences. The nascent strands of replicating molecules form an arc separated from parental and nonreplicating strands. The closer a probe is to its replication origin or to the origin-proximal end of its restriction fragment, the shorter the nascent strands that are detected by the probe. The use of multiple probes allows determination of directions of replication fork movement, as well as locations of origins and termini. In this study, we used simian virus 40 as a model to demonstrate the feasibility of the method, and we discuss its applicability to other systems.

scholarly journals Two-dimensional gel electrophoretic method for mapping DNA replicons.

1988 ◽  
Vol 8 (4) ◽  
pp. 1408-1413 ◽  
Author(s):  
K A Nawotka ◽  
J A Huberman
Keyword(s):  
Gel Electrophoresis ◽  
Replication Origin ◽  
Replication Fork ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Two Dimensional ◽  
Agarose Gel Electrophoresis ◽  
Electrophoretic Method ◽  
Short Probe

We describe in detail a method which allows determination of the directions of replication fork movement through segments of DNA for which cloned probes are available. The method uses two-dimensional neutral-alkaline agarose gel electrophoresis followed by hybridization with short probe sequences. The nascent strands of replicating molecules form an arc separated from parental and nonreplicating strands. The closer a probe is to its replication origin or to the origin-proximal end of its restriction fragment, the shorter the nascent strands that are detected by the probe. The use of multiple probes allows determination of directions of replication fork movement, as well as locations of origins and termini. In this study, we used simian virus 40 as a model to demonstrate the feasibility of the method, and we discuss its applicability to other systems.

Gel Electrophoresis, Isoelectric Focusing, and Localization of Thymidine Kinase in Normal and Simian Virus 40-Infected Monkey Cells

Intervirology ◽  
1973 ◽  
Vol 2 (3) ◽  
pp. 137-151 ◽  
Author(s):  
Saul Kit ◽  
Wai-Choi Leung ◽  
David Trkula ◽  
Del Rose Dubbs ◽  
George Jorgensen
Keyword(s):  
Thymidine Kinase ◽  
Isoelectric Focusing ◽  
Gel Electrophoresis ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Infected Monkey

Method of mapping DNA replication origins

1985 ◽  
Vol 5 (1) ◽  
pp. 85-92
Author(s):  
L D Spotila ◽  
J A Huberman
Keyword(s):  
Dna Replication ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Replication Forks ◽  
Replication Origins ◽  
Chromosomal Dna ◽  
Dna Restriction Fragments ◽  
Dna Restriction ◽  
Dna Replication Origins

We have developed a method which allows determination of the direction in which replication forks move through segments of chromosomal DNA for which cloned probes are available. The method is based on the facts that DNA restriction fragments containing replication forks migrate more slowly through agarose gels than do non-fork-containing fragments and that the extent of retardation of the fork-containing fragments is a function of the extent of replication. The procedure allows the identification of DNA replication origins as sites from which replication forks diverge. In this paper we demonstrate the feasibility of this procedure, with simian virus 40 DNA as a model, and we discuss its applicability to other systems.

scholarly journals Attenuation of late simian virus 40 mRNA synthesis is enhanced by the agnoprotein and is temporally regulated in isolated nuclear systems.

1985 ◽  
Vol 5 (6) ◽  
pp. 1327-1334 ◽  
Author(s):  
N Hay ◽  
Y Aloni
Keyword(s):  
Gel Electrophoresis ◽  
Blot Hybridization ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Wild Type Virus ◽  
Insertion Mutant ◽  
Dot Blot ◽  
Nuclear Systems

Studies were performed to verify the physiological significance of attenuation in the life cycle of simian virus 40 and the role of agnoprotein in this process. For these purposes, nuclei were isolated at various times after infection and incubated in vitro in the presence of [alpha-32P]UTP under the standard conditions which lead to attenuation. Attenuation was evident by the production of a 94-nucleotide attenuator RNA, revealed by gel electrophoresis. In parallel, the synthesis of agnoprotein was studied at various times after infection by labeling the cells for 3 h with [14C]arginine, lysing them, and analyzing the labeled proteins by gel electrophoresis. Both attenuation and the synthesis of agnoprotein were predominant towards the end of the infectious cycle. At earlier times, there was almost no attenuation and no synthesis of agnoprotein. Moreover, there was almost no attenuation even at the latest times after infection in nuclei isolated from cells infected with simian virus 40 deletion mutants that do not synthesize agnoprotein. Finally, analysis by dot blot hybridization showed higher amounts of cytoplasmic viral RNA in cells infected with an agnoprotein gene insertion mutant, delta 79, that does not produce agnoprotein, compared with cells infected with wild-type virus. The present studies indicate that attenuation is temporally regulated and suggest that agnoprotein enhances attenuation in isolated nuclei and that may also enhance it in vivo.

Sign in / Sign up

Export Citation Format

Share Document