NDLI: Transfection and homologous recombination involving single-stranded DNA substrates in mammalian cells and nuclear extracts.

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Transfection and homologous recombination involving single-stranded DNA substrates in mammalian cells and nuclear extracts.

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Transfection and homologous recombination involving single-stranded DNA substrates in mammalian cells and nuclear extracts.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Moore, P. D. Song, K. Y. Ayares, D. Wallace, L. Kucherlapati, R. Rauth, S.
Abstract	We have examined the ability of single-stranded DNA to participate in homologous recombination reactions in mammalian cells and nuclear extracts derived from them. We have inserted a fragment of the neo gene into the single-stranded DNA phage vector M13 mp11. The neo fragment was derived from a deletion derivative of the prokaryotic-eukaryotic shuttle vector pSV2neo. The resulting single-stranded DNA was mixed with a double-stranded deletion derivative of pSV2neo and tested for recombination in human cells, monkey cells, and nuclear extracts obtained from human cells. We were able to obtain recombinant molecules containing wild-type neo genes in all three systems. Examination of the products of recombination indicated that they resulted from correction of the deletion in the double-stranded DNA substrate. We were unable to detect any extensive conversion of single-stranded DNA into its double-stranded counterpart before it participated in the recombination reaction. We have also tested the ability of single-stranded DNA to yield transfectants. When a single-stranded DNA derivative of the herpes simplex virus thymidine kinase (TK) gene was introduced into mouse L-M(TK-) cells, we were able to obtain TK+ colonies. From these results, we conclude that single-stranded DNA can participate in transfection as well as homologous recombination reactions in mammalian cells.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	15
Volume Number	83
Language	English
Publisher	National Academy of Sciences
Publisher Date	1986-09-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	DNA, Single-Stranded Genetics Recombination, Genetic Transfection Animals Cell Nucleus Physiology Cercopithecus Aethiops DNA, Viral Drug Resistance Genetic Vectors Mice Neomycin Pharmacology Thymidine Kinase 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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