Abstract
We established a bicistronic expression system using an encephalomyocarditis virus (EMCV)-derived internal ribosomal entry site (IRES) element to generate stably transformed Drosophila melanogaster Schneider 2 (S2) cells expressing human rotavirus Wa capsid proteins, VP2 and VP6, for the synthesis of VP2/6 double-layered virus-like particle (DVLP). The EMCV-derived IRES permitted bicistronic translation of recombinant VP6. Recombinant VP2 and VP6 were detected in extracellular fractions of stably transformed S2 cells. A wheel-like DVLP (diam ~ 50–55 nm) with short spikes was produced from the extracellular fraction of stably transformed S2 cells. A bicistronic expression system using an EMCV-derived IRES element can thus be used to express two proteins of interest in stably transformed S2 cells. The bi-or tri-cistronic expression of recombinant VP2/6/7 using stably transformed S2 cells can also be used to produce rotavirus VLPs.
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This work was supported by a grant from the Kyung Hee University in 2009 (KHU-20100159).
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Purpose of work A system has been developed for producing capsid proteins from rotavirus, that is a cause of gastroenteritis and diarrhea in infants and young children, with a view to establishing an effective vaccine against this virus.
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Lee, J.M., Chung, H.Y., Kim, K.I. et al. Synthesis of double-layered rotavirus-like particles using internal ribosome entry site vector system in stably-transformed Drosophila melanogaster . Biotechnol Lett 33, 41–46 (2011). https://doi.org/10.1007/s10529-010-0390-x
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DOI: https://doi.org/10.1007/s10529-010-0390-x