In-Yeast Assembly of Coronavirus Infectious cDNA Clones Using a Synthetic Genomics Pipeline.
Methods Mol Biol
; 2203: 167-184, 2020.
Article
in English
| MEDLINE | ID: covidwho-761352
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).
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Saccharomyces cerevisiae
/
DNA, Complementary
/
Coronavirus
/
Genomics
Topics:
Vaccines
Limits:
Animals
Language:
English
Journal:
Methods Mol Biol
Journal subject:
Molecular Biology
Year:
2020
Document Type:
Article
Affiliation country:
978-1-0716-0900-2_13
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