TY  - JOUR
TI  - Gene disruption/knock-in analysis of mONT3: vector construction by employing both in vivo and in vitro recombinations
AU  - Ikeya, Makoto
AU  - Kawada, Masako
AU  - Nakazawa, Yoko
AU  - Sakuragi, Makoto
AU  - Sasai, Noriaki
AU  - Ueno, Morio
AU  - Kiyonari, Hiroshi
AU  - Nakao, Kazuki
AU  - Sasai, Yoshiki
T2  - The International Journal of Developmental Biology
AB  - 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.
PY  - 2005
DO  - 10.1387/ijdb.051975mi
VL  - 49
IS  - 7
SP  - 807
EP  - 823
J2  - Int. J. Dev. Biol.
LA  - en
SN  - 0214-6282
SN  - 1696-3547
UR  - https://ijdb.ehu.eus/article/051975mi
Y2  - 2024/11/24/17:18:34
ER  -