Synthesis and cloning of gene encoding fusion rotaviral VP4-VP6 peptide

Each year, 111 million case of rotavirus diarrhea are reported. There is no specific therapy to treat rotavirus diarrhea, effective contraception should be considered. Considering water and food hygiene appears to have little effect on disease control, vaccination is the only way to reduce death caused by rotavirus diarrhea. In the current study, using modern genetic engineering method, a Polypeptid composed of several antigenic epitopes is made which can be used as a vaccine for rotavirus if its efficiency is cinfirmed. In this study, the prevalence of different species of viruses in Iran was examined. Serotype A was been the most common serotypes. The complete genome sequence was been, antigenic structure of the virus surface proteins were predicted by the of bioinformatics tools including Hex software and server Phyre. Four epitopes of the two virus surface proteins were identified and their genomes were sequenced. Primers were designed by use of software Genrunner in the way that each primer pair formed an epitope. Primers had overlapping areas, SOE-PCR method was used to connect them. In the gel electrophoresis, primers binding and the components growth are clearly seen. Combined fragment was cloned and initial expression was studied in prokaryotic system. The final product was place exactly at the desired place in comparison whit the marker according to molecular weight. Implementing SOE-PCR method in the study, encoding sequence of four epitopes of two rotavirus protein had bind together and new recombined piece was expressed in prokaryotic system. New recombined protein was in the expected place in terms of molecular weight.