title: Engineering infectious cDNAs of coronavirus as bacterial artificial chromosomes.
abstract: The large size of the coronavirus (CoV) genome (around 30 kb) and the instability in bacteria of plasmids carrying CoV replicase sequences represent serious restrictions for the development of CoV infectious clones using reverse genetic systems similar to those used for smaller positive sense RNA viruses. To overcome these problems, several approaches have been established in the last 13 years. Here we describe the engineering of CoV full-length cDNA clones as bacterial artificial chromosomes (BACs), using the Middle East respiratory syndrome CoV (MERS-CoV) as a model.
authors.affiliation: Department of Molecular and Cell Biology, Centro Nacional de Biotecnología (CNB-CSIC), Campus Universidad Autónoma de Madrid, Darwin 3, Cantoblanco, Madrid, 28049, Spain.
authors.forename: Fernando
authors.initials: F
authors.lastname: Almazán
authors.forename: Silvia
authors.initials: S
authors.lastname: Márquez-Jurado
authors.forename: Aitor
authors.initials: A
authors.lastname: Nogales
authors.forename: Luis
authors.initials: L
authors.lastname: Enjuanes
mesh_terms: D000818:Animals; D002460:Cell Line; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D017934:Coronavirus / Q000235:genetics*; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics*; D004926:Escherichia coli; D005818:Genetic Engineering; D006801:Humans; D010957:Plasmids / Q000235:genetics / Q000302:isolation & purification; D059386:Reverse Genetics; D014170:Transformation, Genetic
chemical_list: D018076:DNA, Complementary