An in-vitro transcription assay for development of Rotavirus VP7.

ABSTRACT:

Background and objectives

Human rotavirus (RV) is responsible for most cases of acute gastroenteritis in infants, worldwide. Today, in vitro transcription (IVT) assay is widely used to develop efficient RNA for the biological experiments such as gene function analysis and reverse genetics. The aim of this study was to develop optimal full-length transcripts of the VP7 segment, using in vitro transcription assay.

Materials and methods

Special primers were designed in order to synthesize VP7 sequence of sense RNA in the process of IVT using T7 RNA polymerase. RT-PCR was performed using forward and reverse primers, containing T7 promoter sequence and BstUI restriction enzyme site, respectively. In order to synthesize ssRNA VP7, in accordance with the IVT te

SUBMITTER: Jalilian S 

PROVIDER: S-EPMC5719513 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature