Establishment of stable cell line for inducing KAP1 protein expression

Liu, Xiaoyan and Khan, Md. Asaduzzaman and Cheng, Jingliang and Wei, Chunli and Zhang, Lianmei and Fu, Junjiang (2015) Establishment of stable cell line for inducing KAP1 protein expression. Acta Biologica Hungarica, 66 (2). pp. 161-168. ISSN 0236-5383

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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.

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