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Fig. 6. | BMC Biology

Fig. 6.

From: MultiBac: from protein complex structures to synthetic viral nanosystems

Fig. 6.

Minimizing and optimizing the baculoviral genome to create SynBac. a The DNA elements in the wild-type AcMNPV baculoviral genome were classified according to perceived importance for cell culture applications, derived from exhaustive comparative genome analysis and data mining [69], and distributed into fragments (I to VII) for rewiring and condensing, with the aim to eliminate surplus DNA and improve virus performance in the laboratory and in manufacturing. b Experiments with SynBac1.0, a MultiBac-derivative comprising a condensed Fragment I, showed powerful expression of a test protein (marked by arrowhead), indistinguishable from MultiBac based on Coomassie-stained SDS-PAGE. Numbers indicate molecular weight marker sizes (in kDa). T total cell extract, S soluble protein. c Serial passaging of SynBac1.0 expressing a large polygenic test open reading frame (ORF) containing a red fluorescent protein marker (dsRED) evidenced a remarkable increase of SynBac1.0 genome stability compared with MultiBac. More than 70% of infected cells still produce dsRED in fluorescent micrographs, indicating the presence of intact genomes. By contrast, identically overamplified MultiBac virus frequently lost dsRED, resulting in a much lower proportion (<20%) of infected cells producing dsRED

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