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PMID: 1537326 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Molecular complementation of a collagen mutation in mammalian cells using yeast artificial chromosomes.

The EMBO journal ·Vol. 11 ·No. 2 ·1992-02-00 ·Pages 417-22

Strauss WM, Jaenisch R

Abstract

The cloning of large contiguous segments of mammalian DNA in Saccharomyces cerevisiae has become possible with the advent of Yeast Artificial Chromosomes (YACs). We are interested in extending the technique of genetic complementation analysis to the molecular level through the introduction of YACs into mammalian cells and the mammalian germline. We report the successful introduction of homogeneous DNA derived from a 150 kbp YAC spanning the murine Col1a1 locus into murine fibroblasts carrying a mutation at this locus. The YAC DNA was fractionated by pulse field electrophoresis, condensed with polyamines, and introduced into mutant fibroblasts via DNA-lipid micelles. The DNA was integrated as a stable intact unit in 10% of the transfected clones conferring collagen RNA expression to the mutant cells.

Related Genes
Col1a1
MeSH Terms
Animals Chromosomes, Fungal Collagen/genetics Crosses, Genetic DNA/genetics DNA Probes Genetic Complementation Test Mice Muridae Mutation Restriction Mapping Saccharomyces cerevisiae/genetics Transfection
Chemicals
DNA Probes Collagen DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Strauss W M
Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge 02142.
Jaenisch R
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1992-02-00
Pages
417-22
Language
English
Region
England
NLM ID
8208664
PMCID
PMC556470
Subset
IM
Grants
NIGMS NIH HHS · 5 F32 GM13756-02 · United States
NCI NIH HHS · 5 R35 CA44339-05 · United States
NHGRI NIH HHS · HG000198-01 · United States
External Links
PubMed source PMC Full Text View journal details
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