Efficient cloning of tilapia lake virus complementary DNAs using an in vivo strategy in baker's yeast

• Autores: Cueva, Mario D., Villena, Gretty K., Kitazono, Ana A.
• Formato: artículo
• Fecha de Publicación: 2021
• Institución: Consejo Nacional de Ciencia Tecnología e Innovación
• Repositorio: CONCYTEC-Institucional
• Lenguaje: inglés
• OAI Identifier: oai:repositorio.concytec.gob.pe:20.500.12390/2428
• Enlace del recurso: https://hdl.handle.net/20.500.12390/2428; https://doi.org/10.1111/jwas.12776
• Nivel de acceso: acceso abierto
• Materia: viral recombinant proteins; plasmid construction; Saccharomyces cerevisiae in vivo cloning; http://purl.org/pe-repo/ocde/ford#3.04.03

Cloning and protein expression in heterologous systems are very useful tools for the study of viral proteins. In this work, an in vivo cloning strategy was applied using the yeast Saccharomyces cerevisiae, as an efficient and low-cost method to clone several cDNAs from the tilapia lake virus (TiLV). Samples of infected tilapia Oreochromis niloticus tissues were taken and used to isolate their RNA and to obtain and clone the ten viral cDNAs in a shuttle plasmid. The cloning efficiencies range from 5 to 100% but for seven of the cDNAs the values were above 40%, demonstrating the high efficiency of the method. © 2021 The Authors. Journal of the World Aquaculture Society published by Wiley Periodicals LLC on behalf of World Aquaculture Society.