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PMID: 7935385 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Homology dependence of targeted recombination at the Chinese hamster APRT locus.

Molecular and cellular biology ·Vol. 14 ·No. 10 ·1994-10-00 ·Pages 6663-73

Scheerer JB, Adair GM

Abstract

Using simple linear fragments of the Chinese hamster adenine phosphoribosyltransferase (APRT) gene as targeting vectors, we have investigated the homology dependence of targeted recombination at the endogenous APRT locus in Chinese hamster ovary (CHO) cells. We have examined the effects of varying either the overall length of targeting sequence homology or the length of 5' or 3' flanking homology on both the frequency of targeted homologous recombination and the types of recombination events that are obtained. We find an exponential (logarithmic) relationship between length of APRT targeting homology and the frequency of targeted recombination at the CHO APRT locus, with the frequency of targeted recombination dependent upon both the overall length of targeting homology and the length of homology flanking each side of the target gene deletion. Although most of the APRT+ recombinants analyzed reflect simple targeted replacement or conversion of the target gene deletion, a significant fraction appear to have arisen by target gene-templated extension and correction of the targeting fragment sequences. APRT fragments with limited targeting homology flanking one side of the target gene deletion yield proportionately fewer target gene conversion events and proportionately more templated extension and vector correction events than do fragments with more substantial flanking homology.

MeSH Terms
Adenine Phosphoribosyltransferase/genetics Animals CHO Cells Cricetinae Cricetulus/genetics Gene Conversion Gene Deletion Genetic Vectors Mutagenesis, Insertional Nucleic Acid Hybridization Recombination, Genetic Restriction Mapping Selection, Genetic Sequence Homology, Nucleic Acid
Chemicals
Adenine Phosphoribosyltransferase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Scheerer J B
University of Texas M. D. Anderson Cancer Center, Science Park-Research Division, Smithville 78957.
Adair G M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.14.10.6663-6673.1994
Published
1994-10-00
Pages
6663-73
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC359196
Subset
IM
Grants
NCI NIH HHS · CA 28711 · United States
External Links
PubMed source DOI: 10.1128/mcb.14.10.6663-6673.1994 PMC Full Text View journal details
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