title: Identification of mutations in a temperature-sensitive mutant (tsm5) of murine cytomegalovirus using complementary genome sequencing.
abstract: Identification of mutations in mutants derived chemically is a difficult and relatively random process. NimbleGen Comparative Genome Sequencing (CGS) was assessed as an inexpensive, rapid method of identifying mutations in the temperature-sensitive mutant tsm5 of the K181 (Birmingham) variant of murine cytomegalovirus (MCMV). This genome resequencing approach requires an established genome sequence as a reference. Comparison of tsm5 and the K181 (Birmingham) variant with the published K181 (Perth) MCMV genomic sequence revealed a total of 10 synonymous and 15 non-synonymous SNPs in tsm5 and 14 of the latter were confirmed by sequencing. Thus, while CGS cannot be relied upon to identify correctly all mutations it was helpful for identifying a large number of mutations for further investigation that could contribute to the ts phenotype of tsm5.
authors.affiliation: School of Biosciences, University of Birmingham, Edgbaston, Birmingham, United Kingdom.
authors.forename: Olga
authors.initials: O
authors.lastname: Timoshenko
authors.forename: Abdulaziz
authors.initials: A
authors.lastname: Al-Ali
authors.forename: Brian A B
authors.initials: BA
authors.lastname: Martin
authors.forename: Clive
authors.initials: C
authors.lastname: Sweet
mesh_terms: D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D020043:Genes, Essential; D005814:Genes, Viral; D016679:Genome, Viral; D018146:Muromegalovirus / Q000235:genetics*; D020125:Mutation, Missense; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: D004279:DNA, Viral