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Figure 1 | BMC Biotechnology

Figure 1

From: Engineering of N. benthamianaL. plants for production of N-acetylgalactosamine-glycosylated proteins - towards development of a plant-based platform for production of protein therapeutics with mucin type O-glycosylation

Figure 1

Representative molecular analyses of N. benthamiana L. GalNAc-T2 transgenic plants. A - Schematic diagram of T-DNA of pH7WG2:GNT2 plant binary expression vector. Abbreviations: LB, left border; RB, right border; Pnos, nopaline synthase promoter; P35 S, CaMV 35 S promoter; hpt, hygromycin phosphotransferase gene; Tnos, 3' termination signal of nopaline synthase gene; T35 S, 3' termination signal of CaMV 35 S gene; B - PCR screen using 1 μg genomic DNA as a template and primers G1/G2 (see "Materials and Methods") amplifying human GalNAc-T2 gene (1.7 kb); C - RT-PCR using 0.5 μg total RNA as a template and primers G3/G4 (see "Materials and Methods) that amplify a 0.96 kb fragment of the GalNAc-T2 transcript; D - Western blot of total hydrophobic proteins probed with antibodies raised against human GalNAc-T2; E - The same protein samples separated by SDS-PAGE (4-20% gradient gel) after staining with Coomassie Brilliant Blue R-250; Abbreviations used: C - control plant; 1,2 - putative transgenic plants; M- markers: Promega 1 kb DNA ladder (B, C), BioRad Kaleidoscope prestained standards (D, E).

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BMC Biotechnology

ISSN: 1472-6750

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