NDLI: DNA supercoiling of recombinant plasmids in mammalian cells.

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DNA supercoiling of recombinant plasmids in mammalian cells.

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B- to Z-DNA transition probed by oligonucleotides containing methylphosphonates.

Human acid beta-glucosidase: isolation and amino acid sequence of a peptide containing the catalytic site.

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Structure and expression of the murine N-myc gene.

Resistance to human respiratory syncytial virus (RSV) infection induced by immunization of cotton rats with a recombinant vaccinia virus expressing the RSV G glycoprotein.

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DNA supercoiling of recombinant plasmids in mammalian cells.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Shen, C. K. Hu, W. S.
Abstract	We have used chloroquine/agarose gel electrophoresis and a blot-hybridization technique to study the modulation of superhelicity of extrachromosomal DNA in mammalian cells. The high sensitivity of the procedure has allowed us to measure the change in the specific linking difference or superhelical density (sigma) of a plasmid, psvo alpha 1p3d, after its introduction into COS-7 cells by DNA transfection. Because the molecular weight of psvo alpha 1p3d is approximately the same as that of simian virus 40 (SV40) DNA, the latter can be used as a standard for estimating the average linking difference or number of superhelical turns (tau) of psvo alpha 1p3d after separation of the different supercoiled species on chloroquine/agarose gels. It was found that transfection of monkey cells with either fully supercoiled psvo alpha 1p3d isolated from bacteria (tau = -27 +/- 1, sigma congruent to -0.051) or its relaxed form after treatment with DNA topoisomerase I yields psvo alpha 1p3d samples of the same tau and sigma values of -20 +/- 1 and -0.038, respectively. The difference between the tau values of psvo alpha 1p3d and SV40 in COS-7 cells, in which both plasmids undergo rounds of replication, corresponds to an average difference of 5 +/- 1 superhelical turns. Plasmid psvo alpha 1p3d remains at this lower level of superhelicity for at least 72 hr. The distribution in linking numbers of the topoisomers of psvo alpha 1p3d isolated from transfected COS cells is also more heterogeneous than that of SV40 DNA. These results suggest that the regulation of DNA supercoiling and chromatin assembly may be closely associated with specific DNA sequences. The approach presented here should have a wide application in the study of the regulation and functional role(s) of DNA supercoiling of plasmids in mammalian cells.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	6
Volume Number	83
Language	English
Publisher	National Academy of Sciences
Publisher Date	1986-04-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	DNA, Recombinant DNA, Superhelical Fibroblasts Genetic Vectors Plasmids Animals Cell Line Cercopithecus Aethiops Chloroquine Chromatin Ultrastructure DNA Replication DNA, Viral Electrophoresis, Agar Gel Kidney Nucleic Acid Conformation Simian Virus 40 Genetics Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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