Efficient cloning of tilapia lake virus complementary DNAs using an in vivo strategy in baker's yeast
Open Access
- 3 May 2021
- journal article
- research article
- Published by Wiley in Journal of the World Aquaculture Society
- Vol. 52 (6) , 1209-1220
Abstract
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.Keywords
This publication has 34 references indexed in Scilit:
- Chromatographically-purified capsid proteins of red-spotted grouper nervous necrosis virus expressed in Saccharomyces cerevisiae form virus-like particlesProtein Expression and Purification, 2013
- Bacterial expression systems for recombinant protein production: E. coli and beyondBiotechnology Advances, 2012
- Optimized protocols and plasmids for in vivo cloning in yeastGene, 2011
- Cell Biology of Homologous Recombination in Yeast2011
- Recombinant protein-based viral disease diagnostics in veterinary medicineExpert Review of Molecular Diagnostics, 2010
- Enzymatic assembly of DNA molecules up to several hundred kilobasesNature Methods, 2009
- Saccharomyces cerevisiae: a versatile eukaryotic system in virologyMicrobial Cell Factories, 2007
- Antigenicity of infectious pancreatic necrosis virus VP2 subviral particles expressed in yeastVaccine, 2007
- Highly Efficient Yeast-Based in Vivo DNA Cloning of Multiple DNA Fragments and the Simultaneous Construction of Yeast/ Escherichia Coli Shuttle VectorsBioTechniques, 2006
- Recombination-mediated PCR-directed plasmid construction in vivo in yeastNucleic Acids Research, 1997