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

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)....

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Bibliographic Details
Published inJournal of the World Aquaculture Society Vol. 52; no. 6; pp. 1209 - 1220
Main Authors Cueva, Mario D., Villena, Gretty K., Kitazono, Ana A.
Format Journal Article
LanguageEnglish
Published Hoboken, USA Wiley Subscription Services, Inc 01.12.2021
John Wiley & Sons, Inc
Subjects
Baking yeast
Cloning
Freshwater fishes
In vivo methods and tests
Lakes
Marine fishes
plasmid construction
Plasmids
Ponds
Proteins
Saccharomyces cerevisiae in vivo cloning
Tilapia
viral recombinant proteins
Viruses
Yeast
Yeasts
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Summary: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.
ISSN:0893-8849
1749-7345
DOI:10.1111/jwas.12776