An Insect Nidovirus Emerging from a Primary Tropical Rainforest

Zirkel, Florian; Kurth, Andreas; Quan, Phenix-Lan; Briese, Thomas; Ellerbrok, Heinz; Pauli, Georg; Leendertz, Fabian H.; Lipkin, W. Ian; Ziebuhr, John; Drosten, Christian; Junglen, Sandra

The ORF2a probe detected the genome and two additional bands, one corresponding to a subgenomic RNA (sgRNA) starting upstream of ORF2a, the other within ORF2a (Fig. 4A) . No band corresponding to a separate sgRNA for the predicted ORF2b was observed. A prominent band of 1.8 kb was seen with all probes in and downstream of ORF3a, suggesting an sgRNA starting upstream of ORF3a. A minor band of 1.4 kb was confirmed with the probe placed in ORF3b and all downstream probes. Two additional minor bands of ca. 1.2 and 1.0 kb were seen with the ORF4 probe and the 3=-end probe. According to estimated molecular weights, these corresponded to sgRNAs starting ca. 300 and 500 nt upstream of the initiation codon of ORF4 ( 19399 AUG). Functional studies are required to determine these positions more precisely, as size estimates are based on RNA gel electrophoresis only.