Abstract: The viral total RNA was extracted from dead larvae of the tea looper, Ectropis oblique infected by E. oblique picorna-like virus (EoPV). Five pairs of primers were designed according to the published sequence of EoPV to amplify 5 overlapping fragments covering EoPV genome. These fragments were subcloned to pMD18-T vector and assembled. The full-length cDNA of p-EoPV was cloned to pET-28a and proved by sequencing and restrictive digestion. Comparing to the published sequence, this clone possessed an 8-amino acid mutation and a 1-amino acid deletion. This study represents the first step towards understanding the mechanism of EoPV replication.  

Key words: Ectropis oblique picorna-like virus, full-length cDNA clone, Iflaviridae, reverse genetics manipulation technique