Zhan X, Lee M, Abenes G, Von Reis I, Kittinunvorakoon C, Ross-Macdonald P, Snyder M, Liu F
Program in Infectious Diseases and Immunity, School of Public Health, University of California, 140 Warren Hall, Berkeley, California, 94720, USA.
Virology. 2000 Jan 20;266(2):264-74. doi: 10.1006/viro.1999.0089.
A transposon derived from Escherichia coli Tn3 was introduced into the genome of murine cytomegalovirus (MCMV) to generate a pool of viral mutants. We analyzed three of the constructed recombinant viruses that contained the transposon within the M25, M27, and m155 open reading frames. Our studies provide the first direct evidence to suggest that M25 and M27 are not essential for viral replication in mouse NIH 3T3 cells. Studies in cultured cells and Balb/c mice indicated that the transposon insertion is stable during viral propagation both in vitro and in vivo. Moreover the virus that contained the insertion mutation in M25 exhibited a titer similar to that of the wild-type virus in the salivary glands, lungs, livers, spleens, and kidneys of the Balb/c mice that were intraperitoneally infected with these viruses. These results suggest that M25 is dispensable for viral growth in these organs and the presence of the transposon sequence in the viral genome does not significantly affect viral replication in vivo. The Tn3-based system can be used as a mutagenesis approach for studying the function of MCMV genes in both tissue culture and in animals.
将源自大肠杆菌Tn3的转座子引入鼠巨细胞病毒（MCMV）基因组，以产生一组病毒突变体。我们分析了构建的三种重组病毒，它们在M25、M27和m155开放阅读框内含有转座子。我们的研究提供了首个直接证据，表明M25和M27对病毒在小鼠NIH 3T3细胞中的复制并非必不可少。在培养细胞和Balb/c小鼠中的研究表明，转座子插入在病毒体外和体内传播过程中是稳定的。此外，在M25中含有插入突变的病毒，在腹腔感染这些病毒的Balb/c小鼠的唾液腺、肺、肝、脾和肾中的滴度与野生型病毒相似。这些结果表明，M25对于这些器官中的病毒生长是可有可无的，并且病毒基因组中转座子序列的存在不会显著影响病毒在体内的复制。基于Tn3的系统可作为一种诱变方法，用于研究MCMV基因在组织培养和动物中的功能。
