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PMID: 7604039 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Capturing genes encoding membrane and secreted proteins important for mouse development.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 92 ·No. 14 ·1995-07-03 ·Pages 6592-6

Skarnes WC, Moss JE, Hurtley SM, Beddington RS

Abstract

A strategy based on the gene trap was developed to prescreen mouse embryonic stem cells for insertional mutations in genes encoding secreted and membrane-spanning proteins. The "secretory trap" relies on capturing the N-terminal signal sequence of an endogenous gene to generate an active beta-galactosidase fusion protein. Insertions were found in a cadherin gene, an unc6-related laminin (netrin) gene, the sek receptor tyrosine kinase gene, and genes encoding two receptor-linked protein-tyrosine phosphatases, LAR and PTP kappa. Analysis of homozygous mice carrying insertions in LAR and PTP kappa showed that both genes were effectively disrupted, but neither was essential for normal embryonic development.

MeSH Terms
Animals Base Sequence Cadherins/biosynthesis Cell Line Cloning, Molecular Cricetinae DNA Primers Embryo, Mammalian Embryonic and Fetal Development Fetal Proteins/biosynthesis Kidney Laminin/biosynthesis Mice Molecular Sequence Data Mutagenesis, Insertional Oligodeoxyribonucleotides Polymerase Chain Reaction Protein Tyrosine Phosphatases/biosynthesis Protein-Tyrosine Kinases/biosynthesis Receptor Protein-Tyrosine Kinases/biosynthesis Receptor, EphA4 Recombinant Fusion Proteins/biosynthesis Stem Cells/cytology,physiology Transfection beta-Galactosidase/biosynthesis
Chemicals
Cadherins DNA Primers Fetal Proteins Laminin Oligodeoxyribonucleotides Recombinant Fusion Proteins Protein-Tyrosine Kinases Receptor Protein-Tyrosine Kinases Receptor, EphA4 Protein Tyrosine Phosphatases beta-Galactosidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Skarnes W C
Biotechnology and Biological Sciences Research Council, Centre for Genome Research, University of Edinburgh, United Kingdom.
Moss J E
Hurtley S M
Beddington R S
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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.92.14.6592
Published
1995-07-03
Pages
6592-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC41564
Subset
IM
Grants
Wellcome Trust · United Kingdom
Databases
GENBANK
U23505, U23536
External Links
PubMed source DOI: 10.1073/pnas.92.14.6592 PMC Full Text View journal details
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