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Establishment and Characterization of Inducible Cell Line with Stable Expression of Spata3
Jun-Jiang Fu1,2,3*, Wei-Min Zeng1*, Yue-Hong Li4, Ming-Ri Guo1
1Department of Biochemistry, School of Biological Science and Technology, Central South University, Changsha 410013, China; 2Institute of Human Reproductive and Stem Cell Engineering, Central South University, Changsha 410078, China;
Abstract: Spata3 is a testis-specific expression gene and may play a role in spermatogenesis or spermatogenesis cell apoptosis. To further investigate the spata3 function， we successfully established the cell line which can stably express Flp-InTMT-RExTM-spata3 under the induction of tetractcline or doxycline by cloning the spata3 ORF into our modified pcDNA5/FRT/TO expression vector， with the Flp-InTMT-RExTM-293 cell line as a host line. This cell line has 2×FLAG tag and 2×His tag in the 3'- prime of spata3 and the full-length protein of spata3 can therefore be easily detected by Western blotting with FLAG tag and/or His tag antibody. This procedure is not only very useful for spata3 functional study， but also a useful reference for other protein complex(s) purification and function identification by using highly specific anti-FLAG antibody.
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