Synthesis of double-layered rotavirus-like particles using internal ribosome entry site vector system in stably-transformed Drosophila melanogaster.
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.
Lee
Jong Min
JM
Department of Orthopaedics, Dongguk University Ilsan Hospital, Goyang, 814-820, Korea.
Chung
Ha Young
HY
Kim
Kyung Il
KI
Yoo
Ki Hyun
KH
Hwang-Bo
Jeon
J
Chung
In Sik
IS
Park
Jong-Hwa
JH
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004331:Drosophila melanogaster; D004686:Encephalomyocarditis virus / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014773:Virology / Q000379:methods; D022701:Virosomes / Q000235:genetics / Q000378:metabolism*
D000956:Antigens, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins; C442545:VP2 protein, Rotavirus; C061332:VP6 protein, Rotavirus; D022701:Virosomes