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PMID: 2734308 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Targeted homologous recombination at the endogenous adenine phosphoribosyltransferase locus in Chinese hamster cells.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 12 ·1989-06-00 ·Pages 4574-8

Adair GM, Nairn RS, Wilson JH, Seidman MM, Brotherman KA, MacKinnon C, Scheerer JB

Abstract

We have developed a system that permits analysis of targeted homologous recombination at an endogenous, chromosomal gene locus in cultured mammalian cells. Using a hemizygous, adenine phosphoribosyltransferase (APRT)-deficient, Chinese hamster ovary (CHO) cell mutant as a transfection recipient, we have demonstrated correction of a nonrevertible deletion mutation by targeted homologous recombination. Transfection with a plasmid carrying a fragment of the APRT gene yielded APRT+ recombinants at a frequency of approximately 4.1 x 10(-7). The ratio of targeted recombination to nontargeted integrations of plasmid sequences was approximately 1:4000. Analysis of 31 independent APRT+ recombinants revealed conversions of the endogenous APRT gene, targeted integration at the APRT locus, and a third class of events in which the plasmid donor APRT fragment was converted to a full-length, functional gene.

MeSH Terms
Adenine Phosphoribosyltransferase/deficiency,genetics Animals Blotting, Southern Cell Line Chromosome Mapping Cricetinae Cricetulus Female Genes Genetic Vectors Nucleic Acid Hybridization Ovary Pentosyltransferases/genetics Recombination, Genetic Transfection
Chemicals
Pentosyltransferases Adenine Phosphoribosyltransferase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Adair G M
University of Texas M. D. Anderson Cancer Center, Science Park-Research Division, Smithville 78957.
Nairn R S
Wilson J H
Seidman M M
Brotherman K A
MacKinnon C
Scheerer J B
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.86.12.4574
Published
1989-06-00
Pages
4574-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC287313
Subset
IM
Grants
NCI NIH HHS · CA28711 · United States
NCI NIH HHS · CA36361 · United States
NCRR NIH HHS · RR5511-25 · United States
External Links
PubMed source DOI: 10.1073/pnas.86.12.4574 PMC Full Text View journal details
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