Endah Puji Septisetyani, Adi Santoso


The most common proteins used for reporter protein is the green fluorescent protein (GFP). It is very convenient to detect the GFP fluorescent by fluorescent microscopy or flowcytometry to monitor the successful transfection. The gfp gene can be introduced into the cells by transfecting of two different plasmid vectors or one vector containing both gfp and the gene of our interest. In this current experiment, we used pEGFP-c1 plasmid to express gfp in CHO-K1 cells. We transfected the CHO-K1 cells by using cationic lipid Lipofectamin 2000. We used this study as a way for predicting our human erythropoietin gene expression study in the CHO-K1 cells. In this study, we showed that expression of GFP decreased after incubation of the cells in selection medium containing G418. Expression of GFP seemed to be stable after about three weeks incubation in selection medium. Recombinant erythropoietin was also detected in the day 20.
Key words: Ctionic lipid, CHO-K1 cells, erythropoietin, G418, green fluorescent protein

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Backliwal, G., Hildinger, M., Chenuet, S., Wulhfard, S., Jesus, M. D., and Wurm, F M., 2008, Rational Vector Design and Multi-pathway Modulation of HEK 293E Cells Yield Recombinant Antibody Titers Exceeding 1g/l by Transient Transfection Under Serum-Free Condition, Nuc. Ac. Res., 36(15), e96.

Bollin, F., Dechavanne, V., and Chevalet, L., 2011, Design Experiment in CHO and HEK Transient Transfection Condition Optimization, Protein Exp. and Purif., 78, 61-68.

Çeltikçi, B., Purali, N., Özkara, H. A., 2010, Establishment of Green Fluorescent Protein Expressing CHO Cells by Stable Transfection Using Activated Dendrimers and G418 Selection. Turk. J. Biochem. 35(4), 340-343.

Chen, R., Greene, E. L., Collinsworth, G., Grewal, J. S., Houghton, O., Zeng, H., Garnovskaya, M., Paul, R. V., 1999, Raymond, J. R., Enrichment of Transiently Transfected Mesangial Cells by Cell Sorting after Cotransfection with GFP. Am. J. Physiol. Renal. Physiol. 276, F777-785.

Cheng, L., Du, C., Murray, D., Tong, X., Zhang, Y. A., Chen, B. P., Hawley, R. G., 1997, A GFP Reporter System to Assess Gene Transfer and Expression in Human Hematopoietic Progenitor Cells. Gene Therapy 4, 1013–1022.

Cherng, J. Y., Schuurmans-Nieuwenbroek, N.M., Jiskoot, W., Talsma, H., Zuidam,

N. J., Hennink, W. E., Crommelin, D. J., 1999, Effect of DNA Topology on the Transfection Efficiency of Poly((2dimethylamino)ethyl methacrylate)plasmid Complexes. J. Con. Rel. 60, 343–353.

Gholamin, M., Moaven, O., Farshchian, M., Rajabi-Mashhadi, M. T., Mahmoudi, M.,

Sankian, M., Sazgarnia, A., Ghahraman, M., Abbaszadegan, M. R., 2010, Highly Efficient Transfection of Dendritic Cells Derived from Esophageal Squamous Cell Carcinoma Patient: Optimization with Green Fluorescent Protein and Validation with Tumor RNA as a Tool

for Immuno-genetherapy. Iranian Journal of Biotechnology 8(2), 121-126.

Girard, P., Porte, L., Berta, T., Jordan, M., Wurm, F. M., 2001, Calcium phosphate

transfection optimization for serum-free suspension culture. Cytotechnology 35, 175180.

Hanson, D. A., Ziegler, S. F., 2004, Fusion of Green Fluorescent Protein to the Cterminus of Granulysin Alters Its Intracellular Localization in Comparison to the Native Molecule. J.Neg. Res. inm BioMedicine 3:2.

Huang, H., Hsing, H., Lai, T., Chen, Y., Lee, T., Chan, H., Lyu, P., Wu, C., Lu, Y., Lin, S.,

Lin, C., Lai, C., Chang, H., Chou, H., Chan, H., 2010, Trypsin-Induced Proteome Alteration during Cell Subculture in Mammalian Cells. J. Biom.Scie. 17(36).

Hunt, M. A., Currie, M. J., Robinson, B. A., Dachs, G. U., 2010, Optimizing Transfection of Primary Human Umbilical Vein Endothelial Cells Using Commercially Available Chemical

Transfection Reagents. J. Biomol. Tech. 21(2), 66-72.

Jones, M. L., Seldon, T., Smede, M., Linville, A., Chin, D. Y., Barnard, R., Mahler, S.

M., Munster, D., Hart, D., Gray, P. P., Munro, T. P., 2010, A Method for Rapid,

Ligation-Independent Reformatting of Recombinant Monoclonal Antibodies.

Journal of Immunological Methods 354, 85-90.

Jordan, M., Schallhorn, A., and Wurm, F. M., 1996, Transfecting Mammalian Cells:

Optimization of Critical Parameter Affecting Calcium-Phosphate Precipitate

Formation. Nuc. Ac. Res., 24(4), 596-601.

Leader, B., Baca, Q. J., Golan, D. E., 2008, Protein Therapeutics: a Summary and

Pharmacological Classification. Nature Reviews Drug Discovery 7, 21-39.

Loignon, M., Perret, S., Kelly, J., Boulais, D.,

Cass, B., Bisson, L., Afkhamizarreh, F., and Durocher, Y., 2008, Stable High

Volumetric Production of Glycosylated Human Recombinant IFNalpha2b in

HEK293 Cells, BMC Biotechnology, 8, 65.

Mariati, Ho, S. C. L., Yap, M. G. S., and Yang, Y., 2010, Evaluating PostTranscriptional

Regulatory Elements for Enhancing Transient Gene Expression Levels in CHO K1 and HEK293 Cells, Protein Expr. Purif., 69, 9-15.

Misteli, T., Spector, D. L., 1997, Applications of the Green Fluorescent Protein in Cell Biology and Biotechnology. Nature Biotechnology 15, 961-964.

Rauth, S., Kucherlarapati, R. S., 1984, Expression of DNA transferred into mammalian cells. J. Biosci. 6(4), 543-567.

Stuchbury, G., Munch, G., 2010, Optimizing the Generation of Stable Neuronal Cell

Lines via Pretransfection Restriction Enzyme Digestion of Plasmid DNA. Cytotechnology 62(3), 189-194.

Suzuki, J., Fukuda, M., Kawata, S., Maruoka, M., Kubo, Y., Takeya, T., and Shishido,

T., 2006, A Rapid Protein Expression and Purification System Using Chinese Hamster Ovary Cells Expressing etrovirus Receptor, Journal Biotechnology, 126, 463-474.

Tsien, R. Y., 1998, The Green Fluorescent Protein. Annu. Rev. Biochem. 67, 509-544.



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