Authors:
Xiaoyan Liu Luzhou Medical College, 3-319, Zhongshan road, Luzhou, Sichuan 646000, P. R. China

Search for other papers by Xiaoyan Liu in
Current site
Google Scholar
PubMed
Close
,
Md. Asaduzzaman Khan Luzhou Medical College, 3-319, Zhongshan road, Luzhou, Sichuan 646000, P. R. China

Search for other papers by Md. Asaduzzaman Khan in
Current site
Google Scholar
PubMed
Close
,
Jingliang Cheng Luzhou Medical College, 3-319, Zhongshan road, Luzhou, Sichuan 646000, P. R. China

Search for other papers by Jingliang Cheng in
Current site
Google Scholar
PubMed
Close
,
Chunli Wei Luzhou Medical College, 3-319, Zhongshan road, Luzhou, Sichuan 646000, P. R. China

Search for other papers by Chunli Wei in
Current site
Google Scholar
PubMed
Close
,
Lianmei Zhang Luzhou Medical College, 3-319, Zhongshan road, Luzhou, Sichuan 646000, P. R. China

Search for other papers by Lianmei Zhang in
Current site
Google Scholar
PubMed
Close
, and
Junjiang Fu Luzhou Medical College, 3-319, Zhongshan road, Luzhou, Sichuan 646000, P. R. China

Search for other papers by Junjiang Fu in
Current site
Google Scholar
PubMed
Close
Restricted access

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.

  • 1.

    Andersson, S., Davis, D. L., Dahlbäck, H., Jörnvall, H., Russell, D. W. (1989) Cloning, structure, and expression of the mitochondrial cytochrome P-450 sterol 26-hydroxylase, a bile acid biosynthetic enzyme. J. Biol. Chem. 264, 82228229.

    • Search Google Scholar
    • Export Citation
  • 2.

    Boshart, M., Weber, F., Jahn, G., Dorsch-Häsler, K., Fleckenstein, B., Schaffner, W. (1985) A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus. Cell 41, 521530.

    • Search Google Scholar
    • Export Citation
  • 3.

    Fu, J. J. (2012) Short Protocols in Medical Molecular Biology. China Medical Science Press, Beijing.

  • 4.

    Fu, J., Zhang, L., He, T., Xiao, X., Liu, X., Wang, L., Yang, L., Yang, M., Zhang, T., Chen, R., Xu, J. (2012) TWIST represses estrogen receptor-alpha expression by recruiting the NuRD protein complex in breast cancer cells. Int. J. Biol. Sci. 8, 522532.

    • Search Google Scholar
    • Export Citation
  • 5.

    Fu, J., Qin, L., He, T., Hong, J., Wong, J., Liao, L., Xu, J. (2011) The TWIST/Mi2/NuRD protein complex and its essential role in cancer metastasis. Cell Res. 21, 275289.

    • Search Google Scholar
    • Export Citation
  • 6.

    Fu, J. J., Zeng, W. M., Li, Y. H., Guo, M. R. (2008) Establishment and characterization of inducible cell line with stable expression of Spata3. Chin. J. Cell Biol. 30, 247250.

    • Search Google Scholar
    • Export Citation
  • 7.

    Gritz, L., Davies, J. (1983) Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae. Gene 25, 179188.

    • Search Google Scholar
    • Export Citation
  • 8.

    Iyengar, S., Farnham, P. J. (2011) KAP1 protein: an enigmatic master regulator of the genome. J. Biol. Chem. 286, 2626726276.

  • 9.

    Iyengar, S., Ivanov, A. V., Jin, V. X., Rauscher, F. J. 3rd, Farnham, P. J. (2011) Functional analysis of KAP1 genomic recruitment. Mol. Cell Biol. 31, 18331847.

    • Search Google Scholar
    • Export Citation
  • 10.

    Jakobsson, J., Cordero, M. I., Bisaz, R., Groner, A. C., Busskamp, V., Bensadoun, J. C., Cammas, F., Losson, R., Mansuy, I. M., Sandi, C., Trono, D. (2008) KAP-1 mediated epigenetic expression in the forebrain modulates behavioral vulnerability to stress. Neuron 60, 818831.

    • Search Google Scholar
    • Export Citation
  • 11.

    Liu, L., Zhao, E., Li, C., Huang, L., Xiao, L., Cheng, L., Huang, X., Song, Y., Xu, D. (2013) TRIM28, a new molecular marker predicting metastasis and survival in early-stage non-small cell lung cancer. Cancer Epidemiol. 37, 7178.

    • Search Google Scholar
    • Export Citation
  • 12.

    Moosmann, P., Georgiev, O., Le Douarin, B., Bourquin, J. P., Schaffner, W. (1996) Transcriptional repression by RING finger protein TIF1 beta that interacts with the KRAB repressor domain of KOX1. Nucleic Acids Res. 24, 48594867.

    • Search Google Scholar
    • Export Citation
  • 13.

    Reymond, A., Meroni, G., Fantozzi, A., Merla, G., Cairo, S., Luzi, L., Riganelli, D., Zanaria, E., Messali, S., Cainarca, S., Guffanti, A., Minucci, S., Pelicci, P. G., Ballabio, A. (2001) The tripartite motif family identifies cell compartments. EMBO J. 20, 21402151.

    • Search Google Scholar
    • Export Citation
  • 14.

    Silva, F. P., Hamamoto, R., Furukawa, Y., Nakamura, Y. (2006) TIPUH1 encodes a novel KRAB zincfinger protein highly expressed in human hepatocellular carcinoma. Oncogene 25, 50635070.

    • Search Google Scholar
    • Export Citation
  • 15.

    Wang, C., Ivanov, A., Chen, L., Fredericks, W. J., Seto, E., Rauscher, F. J., Chen, J. (2005) MDM2 interaction with nuclear corepressor KAP1 contributes to p53 inactivation. EMBO J. 24, 32793290.

    • Search Google Scholar
    • Export Citation
  • 16.

    Yokoe, T., Toiyama, Y., Okugawa, Y., Tanaka, K., Ohi, M., Inoue, Y., Mohri, Y., Miki, C., Kusunoki, M. (2010) KAP1 is associated with peritoneal carcinomatosis in gastric cancer. Ann. Surg. Oncol. 17, 821828.

    • Search Google Scholar
    • Export Citation
  • Collapse
  • Expand

Editorial Board

    1. Csányi, Vilmos (Göd)
    1. Dudits, Dénes (Szeged)
    1. Falus, András (Budapest)
    1. Fischer, Ernő (Pécs)
    1. Gábriel, Róbert (Pécs)
    1. Gulya, Károly (Szeged)
    1. Gulyás, Balázs (Stockholm)
    1. Hajós, Ferenc (Budapest)
    1. Hámori, József (Budapest)
    1. Heszky, László (Gödöllő)
    1. Hideg, Éva (Szeged)
    1. E. Ito (Sanuki)
    1. Janda, Tibor (Martonvásár)
    1. Kavanaugh, Michael P. (Missoula)
    1. Kása, Péter (Szeged)
    1. Klein, Éva (Stockholm)
    1. Kovács, János (Budapest)
    1. Brigitte Mauch-Mani (Neuchâtel)
    1. Nässel, Dick R. (Stockholm)
    1. Nemcsók, János (Szeged)
    1. Péczely, Péter (Gödöllő)
    1. Roberts, D. F. (Newcastle-upon-Tyne)
    1. Sakharov, Dimitri A. (Moscow)
    1. Singh, Meharvan (Fort Worth)
    1. Sipiczky, Mátyás (Debrecen)
    1. Szeberényi, József (Pécs)
    1. Székely, György (Debrecen)
    1. Tari, Irma (Szeged)
    1. Vágvölgyi, Csaba (Szeged),
    1. L. Zaborszky (Newark)

 

Acta Biologica Hungarica
P.O. Box 35
H-8237 Tihany, Hungary
Phone: (36 87) 448 244 ext. 103
Fax: (36 87) 448 006
E-mail: elekes@tres.blki.hu

Indexing and Abstracting Services:

  • Biological Abstracts
  • BIOSIS Previews
  • CAB Abstracts
  • Chemical Abstracts
  • Current Contents: Agriculture
  • Biology and Environmental Sciences
  • Elsevier BIOBASE
  • Global Health
  • Index Medicus
  • Index Veterinarius
  • Medline
  • Referativnyi Zhurnal
  • Science Citation Index
  • Science Citation Index Expanded (SciSearch)
  • SCOPUS
  • The ISI Alerting Services
  • Zoological Abstracts

 

Acta Biologica Hungarica
Language English
Size  
Year of
Foundation
1950
Publication
Programme
changed title
Volumes
per Year
 
Issues
per Year
 
Founder Magyar Tudományos Akadémia
Founder's
Address
H-1051 Budapest, Hungary, Széchenyi István tér 9.
Publisher Akadémiai Kiadó
Publisher's
Address
H-1117 Budapest, Hungary 1516 Budapest, PO Box 245.
Responsible
Publisher
Chief Executive Officer, Akadémiai Kiadó
ISSN 0236-5383 (Print)
ISSN 1588-256X (Online)