TY - JOUR ID - 248399 TI - Optimization Of PTS2-EGFP Expression In CHO And Vero Cells JO - Cell Journal (Yakhteh) JA - CELLJ LA - en SN - 2228-5806 AU - Ghodratnama, Roozbeh AU - Karbalaii, Khadijeh AU - Ghaedi, Kamran AU - Baharvand, Hossein AU - Nasr-Esfahani, Mohamad Hossien AD - Stem Cells Department, Royan Institute, Isfahan Research Campus AD - Stem Cells Department, Royan Institute Y1 - 2007 PY - 2007 VL - 9 IS - 3 SP - 170 EP - 175 KW - Expression Vector KW - Targeting signal KW - Lipofection KW - Lipoplex DO - N2 - Objective: Reporter gene transfer to mammalian cells receives a great deal of attention due to its importance for molecular biology, embryology and developmental biology studies. Among DNA transfer technologies to eukaryotic cells, lipofection is known as the most widely used because of its easy handling procedure, low cell mortality and the natural pathway it undertakes.Materials and Methods: In this study we have examined the transfectability of two cell types: CHO and Vero cells via Lipofection in four different treatments, with combination of exposure duration, 3 and 6 hrs, and different plasmid DNA concentration, 0.5 and 1μgs. A fusion protein expression vector, pUcD2. PTS2-EGFP was used to direct the EGFP protein to peroxisomes after expression of related cDNA. An SPSS analysis was preformed after counting the positive cells.Results: optimum gene expression was found when using 1 μg DNA treated for three hrs for CHO cells, and 1 μg DNA treated for six hrs for Vero cells.Conclusion: The result suggests that CHO lipofection efficiency is significantly increased by both the DNA concentration and exposure time increment; however, an increase in exposure time has less significant effect on low DNA concentration conditions. The same results have been observed for Vero cells. Optimum expression was obtained with highest DNA concentration. UR - https://www.celljournal.org/article_248399.html L1 - https://www.celljournal.org/article_248399_08baa1df31ef80fc20cc60aa0d514ea2.pdf ER -