INDONESIAN JOURNAL OF PHARMACY
Vol 24 No 2, 2013

SELECTION OF pEGFP-c1-TRANSFECTED-CHO-K1 CELLS BY G418 DECREASED THE EXPRESSION OF GREEN FLUORESCENT PROTEIN

Endah Puji Septisetyani (Research Center for Biotechnology Indonesian Institute of Sciences Jl. Raya Bogor KM 46, Cibinong, Bogor, Indonesia 16911)
Adi Santoso (Research Center for Biotechnology Indonesian Institute of Sciences Jl. Raya Bogor KM 46, Cibinong, Bogor, Indonesia 16911)



Article Info

Publish Date
01 Apr 2013

Abstract

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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Journal Info

Abbrev

3

Publisher

Subject

Medicine & Pharmacology

Description

Indonesian Journal of Pharmacy (ISSN-e: 2338-9486, ISSN-p: 2338-9427), formerly Majalah Farmasi Indonesia (ISSN: 0126-1037). The journal had been established in 1972, and online publication was begun in 2008. Since 2012, the journal has been published in English by Faculty of Pharmacy Universitas ...