Construction of the Mini-genome of Duck Hepatitis A Virus
In this study we constructed and identified the mini-genome of Duck Hepatitis A virus (DHAV),for analysis of the functional and structural properties of DHAV un-translated region (UTR).To construct a bicistronic of DHAV mini-genome (pRluc-fLuc),the ORF of DHAV was replaced by firefly luciferase (fLuc) gene through enzyme digestion and the Renilla luciferase (Rluc) gene was inserted before 5′ UTR.We also added the hammerhead ribozyme ahead Rluc gene and the hepatitis delta virus ribozyme behind 3′UTR.We transfected the pRluc-fLuc into DF-1 cells,the fLuc gene expression could be detected at 8 hours post transfected (h.p.t.) and with the levels peaking was at 24 h.p.t.The results showed that the DHAV mini-genome was constructed successfully and can be used for explore the role of UTR in virus transcription and translation.
duck hepatitis a virusmini-genomeun-translated region
国家科技期刊平台
NSTL
万方数据
维普
