Enter your keyword

Construction, in silico analysis, and in vitro expression of DNA vaccine candidate encoding human rotavirus capsid protein VP6

Sanawati A.a,b, Barlian A.a, Tan M.I.a

a School of Life Sciences and Technology, Bandung Institute of Technology, Bandung, Indonesia
b Research and Product Development Matrix, Bandung, 40161, Indonesia

Abstract

© OmicsVista Group, All rights reserved.Rotavirus is common cause of severe diarrhea which responsible for 200,000 deaths of children per year. Several live attenuated rotavirus vaccines have been developed and licensed, but it is important to develop new generation of rotavirus vaccine with better safety. In this research, we developed rotavirus DNA vaccine candidate encoding immunogenic VP6 protein from human rotavirus RV4 (HRV-RV4 VP6). VP6 gene was amplified by reverse transcription polymerase chain reaction (RT-PCR) and inserted into pVAX1 vector. The resultant recombinant plasmid, pVAX1-VP6, was analyzed and transfected into mammalian cells. The results revealed that HRV-RV4 VP6 gene, encoding a protein of 397 amino acids, has high homology to VP6 protein of human rotavirus G1P[8]. Three point mutations were found (T246I, H316Y, and I385V) and predicted did not affect the three-dimensional (3D) structure of the HRV-RV4 VP6 protein. However, epitope analysis showed that these mutations might increase the antigenicity of HRV-RV4 VP6. Expression results confirmed that VP6 was expressed since 24 hours after transfection in Vero cells. This study suggested that pVAX1-VP6 can be further developed as rotavirus DNA vaccine candidate because it contained potential epitopes and could be expressed in mammalian cells.

Author keywords

Indexed keywords

DNA vaccine,Epitope,Rotavirus,VP6

Funding details

DOI