Identification of mutations in a temperature‐sensitive mutant ( tsm 5) of murine cytomegalovirus using complementary genome sequencing

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.