Preparation And Expression Analysis Of TRE-C-imp13 Transgenic Mice That Based On Tet System

Since DNA replication and protein expression take part in the different compartment of the eukaryotic cells,nucleocytoplasmic trafficking plays a role in the cell activities.Importin-βfamily members act as nuclear transport receptors in most nuclear transport activities.Importin 13(imp13) is a member of Importin-βfamily, and is found to be the only one bidirectional transport receptor in mammals,which not only functions as an importin,but also an exportin.Several cargoes of importin 13 have been identified:Rbm8,Ubc9,Pax6,the NF-YB/NF-YC dimmer,the gulcocorticoid receptor,myopodin,negative cofactor 2,CHRAC-15/17, p12/CHRAC-17 and eIF-1A.Most of these are essential factors during the development and cell life.The expression or distribution disorders of importin 13 may influence its cargoes to play the role in appropriate time and space.In 2004,Tao et al found that in Hela cell,the C-terminal fragment of rat importin 13 is confined primarily to cytoplasm,while the full-length protein is distributed to both the nucleus and the cytoplasm.Considering this,the newly found transcript of importin 13——TS-imp13 in 2006 may function differently from the full-length importin 13.Here,utilizing microinjection method,we construct a transgenic mouse model of the C-terminal importin 13,aiming at studying the role of importin 13 during mouse development.According to the Tet-on System theory,the transgenic mouse model (TRE-C- imp13) is controlled under the promoter of P_hCMV*-1,the expression of C-imp13 is induced when doxycycline is added to the drinking water of mice.Among the 100 mice born after microinjection,27 mice are tested containing the exogenous C-imp13 gene.Then using RT-PCR and real-time PCR methods,we screen out four founders with relative high expression.These studies result in a newly constructed mice model of C- imp13,making it more convenienct to study the biological function of importin 13 in mice.