Figure 2

EPO cDNA constructs and their expression analysis in CHO cell line. (A) Schematic diagram showing the three EPO expression vectors pcDNA3.1-EPO (without secretory signal, having only mature EPO protein sequence of 495 bp), pcDNA3.1-SS-EPO (with secretory signal fused with mature EPO protein sequence) and pcDNA3.1-K-SS-EPO (having additional six nucleotide of Kozak sequence (GCCACC) before ATG start codon). CMV promoter region of pcDNA 3.1 vector is also shown. (B) mRNA expression analysis showing equal level of expression of all three vectors. 1 μg of the total RNA extracted from CHO cells after 24 hours post transfection was subjected to reverse transcriptase PCR. Lane 1, CHO cells as mock Lane 2, 3 and 4 showing mRNA expression of pcDNA3.1-EPO, pcDNA3.1-SS-EPO and pcDNA3.1-K-SS-EPO respectively. GAPDH is used as internal control. All experiments were performed in duplicate.