title: The high-throughput production of dsRNA against sacbrood virus for use in the honey bee Apis cerana (Hymenoptera: Apidae).
abstract: Sacbrood virus (SBV) is a serious threat to honey bees. Currently, there is no specific drug available for the treatment of SBV that does not affect the quality of the bee product. RNA interference (RNAi) is an important antiviral strategy for disease control. To effectively utilize this technology, the large-scale production and purification of double-stranded RNA (dsRNA) is necessary. Here, a dsRNA-expressing plasmid targeting the VP1 gene of Chinese sacbrood virus (CSBV) was constructed and expressed in Escherichia coli (E. coli) HT115 (DE3). After lysing and ethanol precipitation from E. coli, dsRNA VP1 was purified with an anion exchange chromatography column. Second instar larvae of Apis cerana were fed the purified dsRNA VP1. A significant decrease in larval mortality and the level of expression of the VP1 gene after CSBV infection was demonstrated after the ingestion of dsRNA VP1. This result provides a potential method for the large-scale production of dsRNA to protect A. cerana from CSBV infection.
authors.affiliation: Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Key Laboratory of Integrated Pest Management in Agriculture, Guangdong Entomological Institute, 105 Xingang Road West, Guangzhou, 510260, China.
authors.forename: Jianqing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Key Laboratory of Integrated Pest Management in Agriculture, Guangdong Entomological Institute, 105 Xingang Road West, Guangzhou, 510260, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Key Laboratory of Integrated Pest Management in Agriculture, Guangdong Entomological Institute, 105 Xingang Road West, Guangzhou, 510260, China. hanrc@gdei.gd.cn.
authors.forename: Richou
authors.initials: R
authors.lastname: Han
mesh_terms: D000818:Animals; D001516:Bees / Q000821:virology*; D004926:Escherichia coli / Q000235:genetics; D006927:Hymenoptera / Q000821:virology*; D007304:Insect Viruses / Q000235:genetics*; D007814:Larva / Q000821:virology; D010802:Phylogeny; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics*
chemical_list: D012330:RNA, Double-Stranded
keywords: Apis cerana; RNA interference; Sacbrood virus; dsRNA