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Identification of mutations in a temperature‐sensitive mutant ( tsm 5) of murine cytomegalovirus using complementary genome sequencing
Author(s) -
Timoshenko Olga,
AlAli Abdulaziz,
Martin Brian A.B.,
Sweet Clive
Publication year - 2009
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.21419
Subject(s) - genome , biology , genetics , mutant , mutation , dna sequencing , whole genome sequencing , gene , cytomegalovirus , computational biology , virus , herpesviridae , viral disease
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 tsm 5 of the K181 (Birmingham) variant of murine cytomegalovirus (MCMV). This genome resequencing approach requires an established genome sequence as a reference. Comparison of tsm 5 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 tsm 5 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 tsm 5. J. Med. Virol. 81:511–518, 2009. © 2009 Wiley‐Liss, Inc.