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PMID: 3023904 Published · ppublish English Journal Article

Foreign DNA introduced by calcium phosphate is integrated into repetitive DNA elements of the mouse L cell genome.

Molecular and cellular biology ·Vol. 6 ·No. 5 ·1986-05-00 ·Pages 1787-95

Kato S, Anderson RA, Camerini-Otero RD

Abstract

We investigated the sites of integration of exogenous DNA fragments introduced by DNA-mediated gene transfer. Mouse Ltk- cells were transformed with the herpes simplex virus thymidine kinase gene and pBR322 DNA by the calcium phosphate precipitation method. Some of the integrated exogenous DNA sequences were recovered from the stable tk+ transformants in the form of plasmids that were capable of propagation in bacteria. Four plasmids derived from two cloned cell lines were analyzed in detail by nucleotide sequencing and hybridization techniques. These plasmids contained a total of seven cellular-exogenous DNA junctions. In all cases, there was no sequence homology between the exogenous and cellular DNA sequences adjacent to the joining sites, and no specific exogenous or cellular sequences occurred at the junctions. Rearrangement or deletion of Ltk- DNA was always associated with the integration of exogenous DNA. All of the assignable cellular sequences at the junctions were repetitive sequences. Two of these sequences were from the MIF-1 repetitive sequence family, and a third consisted of a 40-base pair simple copolymer of alternating deoxyadenosine-deoxythymidine. Our results suggest that repetitive sequences are relatively favorable sites for the integration of exogenous DNA.

MeSH Terms
Animals Base Sequence Calcium Phosphates/pharmacology DNA Restriction Enzymes Genes L Cells/enzymology Mice Plasmids Recombination, Genetic Simplexvirus/enzymology,genetics Thymidine Kinase/genetics
Chemicals
Calcium Phosphates Thymidine Kinase DNA Restriction Enzymes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kato S
Anderson R A
Camerini-Otero R D
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.6.5.1787-1795.1986
Published
1986-05-00
Pages
1787-95
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC367708
Subset
IM
Databases
GENBANK
M13105, M13106, M13107, M13108, M13109, M13110, M13265
External Links
PubMed source DOI: 10.1128/mcb.6.5.1787-1795.1986 PMC Full Text View journal details
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