RRC ID 31373
Author Ikeya M, Kawada M, Nakazawa Y, Sakuragi M, Sasai N, Ueno M, Kiyonari H, Nakao K, Sasai Y.
Title Gene disruption/knock-in analysis of mONT3: vector construction by employing both in vivo and in vitro recombinations.
Journal Int J Dev Biol
Abstract We report the isolation, spatial/temporal expression and gene disruption phenotype of the mouse ONT3 (mONT3) gene, which encodes a novel secreted signaling protein belonging to the Olfactomedin/Noelin/Tiarin family. During early embryogenesis, mONT3 is detected in the proximal region of the allantois on embryonic day (E) 7.25, in the lateral plate mesoderm on E 8.0 and in the CNS and heart on E 8.5. The homozygous mutant is born normal and fertile. For the expression pattern and loss-of-function analyses, we have successfully generated the LacZ-knock-in targeting vector directly from BACs carrying mouse genomic fragments by combining in vivo and in vitro recombination techniques. This approach enables rapid and reproducible construction of the fully functional vectors within two weeks without the use of restriction enzyme digestion and ligation, or the use of PCR-amplification of large genomic fragments. In addition, this method is applicable to rapid generation of transgenic vectors, demonstrating its versatility in reverse genetic studies.
Volume 49(7)
Pages 807-23
Published 2005-1-1
DOI 10.1387/ijdb.051975mi
PII 051975mi
PMID 16172977
MeSH Alleles Amino Acid Sequence Animals Biomarkers Chick Embryo Crosses, Genetic Embryo, Mammalian / metabolism Extracellular Matrix Proteins / chemistry Extracellular Matrix Proteins / genetics* Extracellular Matrix Proteins / metabolism Female Gene Deletion* Gene Expression Regulation, Developmental Genetic Vectors / genetics* Genotype Glycoproteins / chemistry Glycoproteins / genetics* Glycoproteins / metabolism Humans Male Mice Mice, Transgenic Molecular Sequence Data Phylogeny Recombination, Genetic / genetics* Sequence Alignment Sequence Homology, Amino Acid Transgenes / genetics*
IF 1.105
Times Cited 32
DNA material nBLUEneo (RDB07039).