title: Establishment of stable cell line for inducing KAP1 protein expression.
abstract: Generation of the stable cell lines is a highly efficient tool in functional studies of certain genes or proteins, where the particular genes or proteins are inducibly expressed. The KRAB-associated protein-1 (KAP1) is an important transcription regulatory protein, which is investigated in several molecular biological studies. In this study, we have aimed to generate a stable cell line for inducing KAP1 expression. The recombinant plasmid pcDNA5/FRT/TO-KAP1 was constructed at first, which was then transfected into Flp-In™T-REx™-HEK293 cells to establish an inducible pcDNA5/FRT/TO-KAP1-HEK293 cell line. The Western blot analysis showed that the protein level of KAP1 is over-expressed in the established stable cell line by doxycycline induction, both dose and time dependently. Thus we have successfully established stable pcDNA5/FRT/TO-KAP1-HEK293 cell line, which can express KAP1 inducibly. This inducible cell line might be very useful for KAP1 functional studies.
authors.affiliation: Key Laboratory of Epigenetics and Oncology, The Research Center for Preclinical Medicine, Luzhou Medical College , 3-319, Zhongshan road, Luzhou, Sichuan 646000 , P. R. China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Liu
authors.forename: Md Asaduzzaman
authors.initials: MA
authors.lastname: Khan
authors.forename: Jingliang
authors.initials: J
authors.lastname: Cheng
authors.forename: Chunli
authors.initials: C
authors.lastname: Wei
authors.forename: Lianmei
authors.initials: L
authors.lastname: Zhang
authors.forename: Junjiang
authors.initials: J
authors.lastname: Fu
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004318:Doxycycline / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D057809:HEK293 Cells; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012097:Repressor Proteins / Q000096:biosynthesis* / Q000235:genetics; D000076183:Tripartite Motif-Containing Protein 28
chemical_list: D000900:Anti-Bacterial Agents; D011994:Recombinant Proteins; D012097:Repressor Proteins; C101116:TRIM28 protein, human; D000076183:Tripartite Motif-Containing Protein 28; D004318:Doxycycline
keywords: KRAB-associated protein-1; inducible cell line; protein expression; recombinant plasmid transfection