[Eukaryotic expression and intracellular distribution of cancer and embryo expression protein 65].
OBJECTIVE
Previous studies show that cancer and embryo expression protein 65 (CEP65) only expresses in various cancer cells and embryo cells. This study was to detect intracellular distribution of CEP65, and make the foundation for further CEP65 function study.


METHODS
Two kinds of eukaryotic expression plasmids, which can separately express full-length CEP65-GST and fluorescent fusion protein CEP65-Red, were constructed, and transfected into COS7 cells by DEAE-Dextran method. Plasmosin and nucleoprotein of COS7 cells were separated. Intracellular distribution of CEP65 was detected by Western blot, and observed under fluorescent microscope.


RESULTS
Western blot confirmed that CEP65 distributed in cytoplasm and nuclei. Fluorescent protein localization showed diffuse distribution of fluorescence in the cells transfected with pcDNA3-Red, and collective fluorescent articles in cytoplasm and nuclei of the cells transfected with pcDNA3-Red-Cep65.


CONCLUSIONS
CEP65 can express efficiently in COS7 cells, and locates in cytoplasm and nuclei. It may be a kind of nucleus-associated protein.
Jin
Geng-Lin
GL
Biochemistry and Molecular Biology Laboratory, Beijing Institute of Cancer Research, School of Oncology, Peking University, Beijing, 100034, P.R.China. jingenglinhappy@yahoo.com.cn
Guo
Jian-Hui
JH
Ma
Hong
H
Meng
Lin
L
D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism*; D019556:COS Cells; D018797:Cell Cycle Proteins; D002467:Cell Nucleus / Q000378:metabolism*; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000378:metabolism*; D003598:Cytoskeletal Proteins; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D000951:Antigens, Neoplasm; D018797:Cell Cycle Proteins; C493903:Cep290 protein, human; D003598:Cytoskeletal Proteins; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins