Fig. 2.

Generation of RNCs in vivo using E. coli expression. After transformation of the NC-containing pLDC-17 vector into E. coli, cell growth to high cell densities in MDG medium was allowed for up to 20 h, 37 °C, 280 rpm. Cells were then harvested and resuspended in M9 expression medium in the absence of a nitrogen source. The nitrogen source was depleted before the addition of ¹⁵N ammonium chloride and expression induced at 25 °C. Cells were harvested 45 min after induction.