Figure 5.

Growth of the transgenic lines of C. reinhardtii secreting Venus and fusion Venus is not affected by the production of the recombinant proteins. (a) Growth curves of the transgenic lines and the parental strain UVM4 under the mixotrophic conditions at 25 °C, under constant illumination at 120 μmol photons m⁻² s⁻¹. Strains were inoculated in TAP media at the OD_750nm value of 0.05. Growth was monitored by cell counting for 7 days. Error bars represent standard errors of at least three independent cultures. There was no statistically significant difference between genotypes based on two‐way ANOVA test (P > 0.05). (b) Time course of Venus‐SP₁₀ and Venus–SP₂₀ secretions in the growth media. Cell line SP04 expressing SSVenus‐(SP)₁₀ (upper panel); cell line SP14 expressing SSVenus‐(SP)₂₀ (lower panel). Samples were taken from shake flasks every 24 h for 7 days. Twenty microlitres of clarified supernatants for each sample were separated on SDS‐PAGE and analysed by Western blot. Lanes 1–7 correspond to samples from 0‐ to 7‐day‐old culture supernatants; molecular size markers are indicated as M. Solid arrow indicates the fusion proteins; open arrows represent non‐glycosylated protein.