Skip to main content
. 2016 Dec 1;11(12):e0167232. doi: 10.1371/journal.pone.0167232

Fig 3. Enhancement of chimeric IgG expression by BGH terminator.

Fig 3

A) Schematic view of the RMCE strategy to replace the native chicken IgM heavy chain terminator with BGH terminator. PhIgG-3 vector contains hIgG1-Fc, a promoter-less blasticidin S resistance gene and the BGH terminator. PhIgG-3 and pCAG-Cre were co-transfected, and the cells in which native terminator is replaced with BGH were selected by the addition of blasticidin S. B) Flow-cytometric analysis with FITC-conjugated anti-hIgG-Fc. The hIgG1 chimeric mAb expressing cells harboring native chicken IgM heavy chain terminator (Native Terminator) and those of which the terminator was replaced with the BGH terminator (BGH Terminator) by RMCE were analyzed. C) Quantitative RT-PCR analysis of the expression levels of membrane-bound hIgG chimeric mAbs. RNA was extracted from cells harboring the native terminator (Native Terminator) and those after RMCE-induced replacement with the BGH terminator (BGH Terminator). The transcript levels were normalized by GAPDH signals. The results are from two independent experiments. The primer annealing sites are indicated by black bars in Fig 3A.