Identification of mutations in a temperature-sensitive mutant (tsm5) of murine cytomegalovirus using complementary genome sequencing

✍ Scribed by Olga Timoshenko; Abdulaziz Al-Ali; Brian A.B. Martin; Clive Sweet

• Publisher: John Wiley and Sons
• Year: 2009
• Tongue: English
• Weight: 514 KB
• Volume: 81
• Category: Article
• ISSN: 0146-6615
• DOI: 10.1002/jmv.21419

✦ Synopsis

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 __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.