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Publication : Gene disruption/knock-in analysis of mONT3: vector construction by employing both in vivo and in vitro recombinations.

First Author  Ikeya M Year  2005
Journal  Int J Dev Biol Volume  49
Issue  7 Pages  807-23
PubMed ID  16172977 Mgi Jnum  J:102991
Mgi Id  MGI:3608304 Doi  10.1387/ijdb.051975mi
Citation  Ikeya M, et al. (2005) Gene disruption/knock-in analysis of mONT3: vector construction by employing both in vivo and in vitro recombinations. Int J Dev Biol 49(7):807-23
abstractText  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.
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