Successful transcription but not translation or assembly of Solenopsis invicta virus 3 in a baculovirus-driven expression system
Keywords:
Solenopsis invicta, unassigned virus, expression, biological controlAbstract
Solenopsis invicta virus 3 (SINV-3) is an unclassified positive stranded RNA virus whose characteristics are amenable to development as a microbial insecticide or as a classical biological control agent for the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae). Bait formulations containing SINV-3 have been produced and were used successfully to transmit the infection and cause significant mortality to fire ant colonies in the laboratory. Unfortunately, there is no available means of propagating infectious SINV-3 particles in vitro, and this has hampered its evaluation and development as a microbial insecticide. In this study, we attempted to utilize a baculovirus expression system as reported by others to produce SINV-3 infectious particles in quantity. A full-length copy of the SINV-3 genome was assembled successfully from 3 overlapping fragments, and a heterologous bacmid-SINV-3 vector (BAC_SINV-3) was produced. The insect cell line Sf21 was transfected with purified high–molecular weight BAC_SINV-3 and supported production of baculovirus expressing full-length SINV-3 transcripts. Despite successful transcription of the SINV-3 genome, evidence that translation was occurring could not be produced. Western analyses for the SINV-3 VP-2 capsid protein were consistently negative.
Resumen
El virus Solenopsis invicta 3 (SINV-3) es un virus ARN de cadena positiva no clasificada, cuyas características son promisorias para el desarrollo de un insecticida microbiano o como agente de control biológico clásico de la hormiga de fuego roja importada, Solenopsis invicta Buren (Hymenoptera: Formicidae). Se han producido y se han utilizado con éxito formulaciones de cebo que contienen SINV-3 para transmitir la infección y causar una mortalidad significativa en las colonias de hormigas de fuego en el laboratorio. Desafortunadamente, no hay medios disponibles para propagar partículas de SINV-3 infecciosas en vitro, lo que ha hecho más dificil su evaluación y su desarrollo como un insecticida microbiano. En este estudio, hemos intentado utilizar un sistema de expresión de baculovirus según lo informado por otros para producir partículas infecciosas de SINV-3 en cantidad. Se produjo con éxito una copia de longitud completa del genoma SINV-3 de 3 fragmentos sobrepuestos, y se produjo un vector heterólogo bácmido-SINV-3 (BAC_SINV-3). La línea celular de insecto, Sf21, se transfectó con BAC_SINV-3 purificada de alto peso molecular que apoyó la producción de baculovirus que expresan lineas enteras de de transcripciones de SINV-3. A pesar de la transcripción con éxito del genoma SINV-3, evidencia de que la traducción estaba ocurriendo no we puede producir. El analisis occidental para la proteína cápside VP-2 de SINV-3 fue consistentemente negativo.
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