| Literature DB >> 32833212 |
Tran Thi Nhu Thao1,2,3, Fabien Labroussaa2,4, Nadine Ebert1,2, Joerg Jores5,6, Volker Thiel7,8.
Abstract
The Escherichia coli and vaccinia virus-based reverse genetics systems have been widely applied for the manipulation and engineering of coronavirus genomes. These systems, however, present several limitations and are sometimes difficult to establish in a timely manner for (re-)emerging viruses. In this chapter, we present a new universal reverse genetics platform for the assembly and engineering of infectious full-length cDNAs using yeast-based transformation-associated recombination cloning. This novel assembly method not only results in stable coronavirus infectious full-length cDNAs cloned in the yeast Saccharomyces cerevisiae but also fosters and accelerates the manipulation of their genomes. Such a platform is widely applicable for the scientific community, as it requires no specific equipment and can be performed in a standard laboratory setting. The protocol described can be easily adapted to virtually all known or emerging coronaviruses, such as Middle East respiratory syndrome coronavirus (MERS-CoV).Entities:
Keywords: Coronavirus; Full-length cDNA clone; Homologous recombination; RNA virus; Reverse genetics; Saccharomyces cerevisiae; Synthetic genomics; Transformation-associated recombination (TAR) cloning
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Year: 2020 PMID： 32833212 DOI： 10.1007/978-1-0716-0900-2_13
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745