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Fusion of the Dhfr/Mtx and IR/MAR Gene Amplification Methods Produces a Rapid and Efficient Method for Stable Recombinant Protein Production

Figure 2

Plasmid structures.

In panels A to K, plasmid sets α to δ are labeled in red; (+) and (−) indicate the presence and absence of IR/MAR, respectively. Double-headed red arrows indicate the co-transfected plasmid set α and γ. Double-headed black arrows indicate the two sets of co-transfected plasmids used to quantify GFP expression by flow cytometry (Figure 3). In each plasmid, blue arrows represent the target genes (d2EGFP, MycL, and MycH) and green arrows represent the drug-resistance genes (Dhfr, BS, and Neo); the IR and MAR are represented by a red dashed bold line and red oval, respectively. The promoters (P-CMV, P-SRα or P-SV40), polyA-addition sequences (TK-pA or SV40-pA), and sequences required for cloning in E. coli (Ori and Amp), are shown in gray. In panel L, the features of the plasmids are summarized. The font colors are the same as those used in panels A to K.

Figure 2

doi: https://doi.org/10.1371/journal.pone.0052990.g002