Development and characterization of a DNA-launched Getah virus infectious clone

Abstract Getah virus (GETV), a neglected and re-emerging mosquito-borne alphavirus, has becoming more serious and posing a potential threat to animal safety and public health. As there is a lack of antivirals and vaccines against GETV, it is necessary to continue the development of tools to further advance our efforts to combat these pathogens, including reverse genetics techniques. Herein, we describe the design and construction of a DNA-launched infectious clone for GETV. The full-length genome of GETV HuN1 strain, flanked by cytomegalovirus immediate-early (CMV) promoter sequence at the 5'-end and the hepatitis delta virus ribozyme along with the bovine growth hormone termination and polyadenylation signal sequences at the 3'-end, was packaged in bacterial artificial chromosome vector to establish a GETV infectious clone pBR322-GETV-HuN1. In parallel, the recombinant reporter viruses carrying the reporter gene EGFP between the E1 gene and the 3' UTR was constructed based on the established CMV-driven cDNA clone. Both in vivo and in vitro experiments have shown that the rescued recombinant virus possesses viral biological activity similar to the parental virus. Taken together, this study develops a concise and efficient GETV infectious cDNA clone and a recombinant virus carrying an EGFP reporter gene. The availability of the GETV infectious clone will facilitate further studies on understanding the molecular mechanisms of GETV virus biology, virulence determinants, molecular pathogenesis, vaccine development and virus-host interaction.