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Figure 2 | Microbial Cell Factories

Figure 2

From: Capsid protein expression and adeno-associated virus like particles assembly in Saccharomyces cerevisiae

Figure 2

Expression of AAV2 Cap and Rep proteins from natural promoters. Transformed RSY12 cells were grown in liquid selective medium until cultures reached different densities determining different growth phases (early → late log) indicated on the top. Equal amounts of total cell lysate, extract 1 and 2 (~50 μg each), obtained from 2 × 108 cells at each phase, were analyzed by Western blot analysis with antibodies indicated on the left: mAb B1 to detect Cap proteins (A and C), mAb 303.9 to detect Rep proteins (B) and mAb Anti-3PGK to detect constitutive yeast protein PGK (Phospho-Glicerate-Kinase). (A): VP3 was the only capsid protein detected in the samples from YEplacp40Cap transformed cells;thehighest level was detected in the mid-log phase extracts. (B): All four Rep proteins were detected in the samples from YEplacRepCap transformed cells; the highest level was obtained in the late-log phase extracts. (C): late-log phase extracts from YEplacRepCap cells were also analyzed for Cap protein expression and compared with late-log phase extracts from YEplacp40Cap cells; only VP3 was detected in both samples. Denatured, 293 T-cell-derived, AAV2 capsids were used as positive control (+ control) for defining VPs (A, C).

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