Maggioncalda J, Mehta A, Fraser N W, Block T M
Department of Microbiology and Immunology, Jefferson Cancer Institute, Thomas Jefferson University, Philadelphia, Pennsylvania 19107-6799.
J Virol. 1994 Dec;68(12):7816-24. doi: 10.1128/JVI.68.12.7816-7824.1994.
A herpes simplex virus type 1 strain 17 mutant with a deletion between genomic nucleotides 118880 and 119250 was constructed and called 17 delta Sty. The deletion removes most of a putative secondary LAT promoter (called LAPII) as well as 370 of the first 449 nucleotides of the proposed 8.5-kb transcript believed to be the precursor of 2.0-kb LAT. 17 delta Sty was shown to produce major 2.0-kb LATs in tissue culture. Moreover, trigeminal nerves from latently infected mice contained an intact 1.45- to 2.0-kb LAT as well as the minor LATs which are recognized by probes specific for regions downstream of the 2.0-kb LAT. Finally, 17 delta Sty reactivated with normal kinetics from the trigeminal ganglia of latently infected mice in the explant cocultivation assay and egressed from tissue culture cells as efficiently as wild-type virus. These results clearly show that the region deleted in 17 delta Sty is dispensable for intact 2-kb LAT production, viral egress in tissue culture, and normal reactivation from latently infected neurons in mice.
构建了一种1型单纯疱疹病毒17株突变体,其基因组核苷酸118880至119250之间存在缺失,命名为17 delta Sty。该缺失去除了一个假定的次要LAT启动子(称为LAPII)的大部分,以及被认为是2.0 kb LAT前体的8.5 kb转录本前449个核苷酸中的370个。17 delta Sty在组织培养中显示产生主要的2.0 kb LAT。此外,来自潜伏感染小鼠的三叉神经含有完整的1.45至2.0 kb LAT以及被2.0 kb LAT下游区域特异性探针识别的次要LAT。最后,在体外共培养试验中,17 delta Sty从潜伏感染小鼠的三叉神经节以正常动力学重新激活,并与野生型病毒一样有效地从组织培养细胞中释放出来。这些结果清楚地表明,17 delta Sty中缺失的区域对于完整的2 kb LAT产生、组织培养中的病毒释放以及小鼠潜伏感染神经元的正常重新激活是可有可无 的。