pXVE-spAAT

Protein Expression Eukaryotic cells - N. benthamiana XVE

Experiment
Protein Expression Eukaryotic cells - N. benthamiana XVE
Product
pXVE-spAAT from Karen A. McDonald, Department of Chemical Engineering and Materi
Manufacturer
Karen A. McDonald, Department of Chemical Engineering and Materi

Protocol tips

Protocol tips
Transgenic N. benthamiana cell suspension cultures for each of the three gene expression systems (35S, XVE and CMViva) expressing recombinant human AAT protein were established and analyzed by ELISA and Western blots for rAAT protein [26,27]. The following cell lines, 35S line #0632, XVE line #6011, and CMViva line #8011, producing high levels of extracellular AAT protein, were evaluated for this study [26].

Publication protocol

Transgenic N. benthamiana cell suspension cultures for each of the three gene expression systems (35S, XVE and CMViva) expressing recombinant human AAT protein were established and analyzed by ELISA and Western blots for rAAT protein [26,27]. The following cell lines, 35S line #0632, XVE line #6011, and CMViva line #8011, producing high levels of extracellular AAT protein, were evaluated for this study [26]. Transgenic N. benthamiana cell suspension cultures were sub-cultured every week by transferring 20 mL of the established suspension cells into 200 mL KCMS medium. The KCMS media consist of 30 g/L sucrose, 4.3 g/L MS (Murashige and Skoog) salt mixture, 0.204 g/L KH2PO4, 0.1 g/L myo-inositol, 10 mg/L thiamine-HCl, 10 mg/L nicotinic acid, 5 mg/L pyridoxine-HCl, 2 mg/L 2,4-D (2,4-dichlorophenoxyacetic acid), and 0.1 mg/L kinetin; the pH 5.8 is adjusted by KOH, in a 1 L flask at 140 rpm and 25 °C.

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