Open AccessEfficient cloning of tilapia lake virus complementary DNAs using an in vivo strategy in baker's yeast
Author(s) -
Cueva Mario D.,
Villena Gretty K.,
Kitazono Ana A.
Publication year - 2021
Publication title -
journal of the world aquaculture society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.655
H-Index - 60
eISSN - 1749-7345
pISSN - 0893-8849
DOI - 10.1111/jwas.12776
Subject(s) - biology , cloning (programming) , oreochromis , clone (java method) , saccharomyces cerevisiae , heterologous , yeast , tilapia , plasmid , heterologous expression , recombinant dna , microbiology and biotechnology , gene , genetics , fish <actinopterygii> , fishery , computer science , programming language
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.