Molecular characterization and expression of human rotavirus recombinant protein VP2, VP6, and VP7 transfected in vero cell
Rahmah L.a,b, Rachman E.G.b, Retroningrum D.S.b, Tan M.I.b
a Rotavirus Vaccine Production PT Biofarma, Bandung, Indonesia
b Institut Teknologi Bandung, Bandung, Indonesia
Abstract
© OmicsVista Group, All rights reserved.Rotavirus is a non-enveloped viruses containing double-stranded RNA genetic material. The 11 ds-RNA encodes 6 structural and 6 nonstructural proteins. Three major structural capsid proteins of human rotavirus RV4 (VP2, VP6, and VP7) had been isolated, cloned and transfected into Vero cell. VP2 (VP2LRV4), VP6 (VP6LRV4), and VP7 (VP7LRV4) was isolated with the size of coding sequence (CDS) 2673 bp, 1194 bp, and 981 bp, respectively. In silico analysis showed that VP2LRV4, VP6LRV4, and VP7LRV4 protein sequences (prediction) comprised essential subdomain and residues to construct triple layer structural capsid protein. Gene transcription and protein expression of VP2LRV4, VP6LRV4 and VP7LRV4 in Vero cell were examined by RT-PCR and immunofluorescence assay (IF). Transcriptional analysis led to the detection of VP6 and VP7 cDNA on day 1 post transfection while VP2 cDNA was expressed on day 3 post transfection. VP6 and VP7 protein expression marked with fluorescence luminescence in Vero cell on day 1 post transfection.
Author keywords
Indexed keywords
Rotavirus,Vero Cell,VP2,VP6,VP7