Characterization of a gene trap insertion into a novel gene, cordon-bleu, expressed in axial structures of the gastrulating mouse embryo
| Autor: | Janet Rossant, John Klingensmith, David P. Hill, Stéphan Gasca |
|---|---|
| Přispěvatelé: | Development and Fetal Health, Mount Sinai Hospital [Toronto, Canada] (MSH), Department of Molecular and Medical Genetics, University of Toronto, Gasca, Stéphan |
| Jazyk: | angličtina |
| Rok vydání: | 1995 |
| Předmět: |
Male
[SDV.GEN] Life Sciences [q-bio]/Genetics MESH: Amino Acid Sequence MESH: Base Sequence Homology (biology) Mice 0302 clinical medicine Genes Reporter MESH: Genetic Vectors MESH: Gene Expression Regulation Developmental MESH: Animals Cloning Molecular Sonic hedgehog [SDV.BDD]Life Sciences [q-bio]/Development Biology MESH: Chimera 0303 health sciences biology Stem Cells Microfilament Proteins Chromosome Mapping Gene Expression Regulation Developmental notochord ES cells Phenotype Lac Operon gene-trap Female MESH: DNA Primers Genetic Vectors Molecular Sequence Data MESH: Lac Opero node Cell Line Embryonic and Fetal Development 03 medical and health sciences Complementary DNA MESH: Gastrula [SDV.BDD] Life Sciences [q-bio]/Development Biology Genetics Animals MESH: Cloning Molecular Amino Acid Sequence gastrulation Gene mouse DNA Primers 030304 developmental biology Reporter gene [SDV.GEN]Life Sciences [q-bio]/Genetics Base Sequence Chimera cDNA library MESH: Genes Reporter Proteins Gastrula Cell Biology Molecular biology MESH: Embryonic and Fetal Development MESH: Cell Line Mice Inbred C57BL Cytoskeletal Proteins Mutagenesis Insertional Open reading frame Fusion transcript biology.protein MESH: Chromosome Mapping MESH: Female 030217 neurology & neurosurgery Developmental Biology |
| Zdroj: | Dev Genet Dev Genet, 1995, 17, pp.141-54. ⟨10.1002/dvg.1020170206⟩ |
| DOI: | 10.1002/dvg.1020170206⟩ |
| Popis: | We have used a gene trap (GT) vector and embryonic stem (ES) cell chimeras to screen for insertions of the lacZ reporter gene into transcription units that are spatially and temporally regulated during early mouse embryogenesis. GT vectors which can act as both a reporter and a mutagen have been previously used to isolate new genes that are essential for mouse development. In this paper we describe a GT insertion which displays a very restricted pattern of expression in the gastrulating embryo. β-Galactosidase activity was first detected at 7.5 days post-coitum (E7.5) in the node region of the embryo and extended to the midline structures at E8.0. At E9.5 expression was restricted to the floor plate, the notochord, the roof of the gut, and the liver anlage. Expression appeared in the somites at E10.0 and later became more widespread. We used rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR) to clone a partial 360 base pair (bp) cDNA representing an endogenous sequence and containing an open reading frame (ORF) fused in frame to the lacZ reporter gene. The sequence showed no homology to any known protein or protein domain. An overlapping 1,200 bp fragment from a wild-type cDNA library was cloned and it detected the same pattern of expression as the reporter gene in E7.5, E8.5, and E9.5 wild-type embryos. It hybridized to a 5.4 kb lacZ fusion transcript and to an endogenous transcript of 6.5 kb. The gene was mapped to chromosome 11 and was named cordon-bleu (cobl). No phenotype was detected in mice homozygous for the insertion. However, the insertion may not cause a complete disruption of the gene function. The pattern of expression of cobl is very similar to that of hepatic nuclear factor 3β (HNF3β) and sonic hedgehog (Shh), both of which are involved in axial patterning. Therefore, the product of the cobl gene may also prove to be an important component of the genetic pathway regulating vertebrate axis formation. © 1995 Wiley-Liss, Inc. |
| Databáze: | OpenAIRE |
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