Isolation of intracellular replicative forms and progeny single strands of DNA from parvovirus KRV in sucrose gradients containing guanidine hydrochloride

Journal Article · Fri Dec 31 23:00:00 EST 1976 · Virology; (United States)

DOI:https://doi.org/10.1016/0042-6822(77)90324-5· OSTI ID:6753556

Lavelle, G; Li, A T

When cells infected with Kilham rat virus (KRV) were incubated in 4 M guanidine-HCl, the single-stranded DNA was completely released from virions of KRV, but double-stranded DNA was not denatured. The three forms of viral DNA released had properties, respectively, of double-stranded replicative-form DNA, single-stranded progeny DNA, and an intermediate DNA species. Sucrose gradients containing 4 M guanidine-HCl were used to isolate these intracellular DNA forms with complete separation from high molecular weight cellular DNA. After a short pulse, radioactivity in low molecular weight DNA was found almost exclusively in molecules having sedimentation properties of viral replicative-form DNA. During chase incubation, this radioactivity progressively shifted to the position of single-stranded progeny DNA.

• Research Organization: Oak Ride National Lab., TN
• OSTI ID: 6753556
• Journal Information: Virology; (United States), Journal Name: Virology; (United States) Vol. 76; ISSN VIRLA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
550700 -- Microbiology
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
CARBOHYDRATES
CARBONIC ACID DERIVATIVES
CENTRIFUGATION
DISACCHARIDES
DNA
GUANIDINES
LABELLED COMPOUNDS
MAMMALS
MICROORGANISMS
MOLECULAR BIOLOGY
MOLECULAR WEIGHT
NUCLEIC ACID DENATURATION
NUCLEIC ACIDS
OLIGOSACCHARIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PARASITES
RATS
RODENTS
SACCHARIDES
SACCHAROSE
SEPARATION PROCESSES
SYNTHESIS
VERTEBRATES
VIRUSES