Cryopreservation ofXenopus transgenic lines

摘要: Xenopus laevis has been widely used for molecular, cellular, and developmental studies. With the development of sperm-mediated transgenic method, it is now possible to study gene function during vertebrate by using this popular model. On other hand, like animal species, labor intensive, maintenance lines expensive. In article, we investigated possibility sperm-cryopreservation as a means preserve frog lines. We demonstrated that cryopreserved sperms are viable but not fertile under our in vitro fertilization (IVF) conditions. However, microinjecting sperm nuclei, successfully regenerated line carrying double promoter transgene construct, where marker encoding green fluorescent protein (GFP) driven γ-crystallin interest, fusion GFP with matrix metalloproteinase stromelysin-3 (ST3-GFP), heat shock-inducible promoter. functional transmission ST3-GFP analyzing phenotype F1 animals after heat-shock induce its expression. Our method thus provides an inexpensive convenient way distribution Furthermore, ease which microinject nuclei compared technically demanding transgenesis procedure variable outcome should facilitate more laboratories use studies vivo. Mol. Reprod. Dev. 67: 65–69, 2004. © 2004 Wiley-Liss, Inc.