Noteborn M H, Verschueren C A, van Ormondt H, van der Eb A J
Laboratory of Molecular Carcinogenesis, Sylvius Laboratories, Leiden University, Leiden, The
Gene. 1998 Nov 26;223(1-2):165-72. doi: 10.1016/s0378-1119(98)00170-x.
Plasmid pCAV/E contains an infectious cloned double-stranded CAV (chicken anemia virus) DNA genome (Noteborn et al., J. Virol. 65 (1991) 3131-3139). We have constructed mutated CAV genomes by introducing mutations into the CAV promoter/enhancer region of pCAV/E. Various mutated CAV strains were functional and had a smaller cytopathogenic effect in chicken T cells than wild-type CAV. In particular, mutations within the '12-bp insert' of the promoter/enhancer region had this effect. PCR and sequence analysis showed that the CAV mutants were stable under cell-culture conditions. Southern-blot analysis showed that all replication DNA intermediates were normally formed by the CAV mutants. All viable mutant CAV strains were able to produce a neutralizing conformational epitope, which implies that they can trigger the required protective immune response. These features make these mutant CAV strains potential candidates for the development of an attenuated CAV vaccine.
质粒pCAV/E包含一个具有感染性的克隆双链CAV（鸡贫血病毒）DNA基因组（Noteborn等人，《病毒学杂志》65 (1991) 3131 - 3139）。我们通过在pCAV/E的CAV启动子/增强子区域引入突变构建了突变的CAV基因组。各种突变的CAV毒株具有功能，并且在鸡T细胞中比野生型CAV具有更小的细胞病变效应。特别地，启动子/增强子区域“12 bp插入序列”内的突变具有这种效应。PCR和序列分析表明，CAV突变体在细胞培养条件下是稳定的。Southern杂交分析表明，所有复制DNA中间体均由CAV突变体正常形成。所有存活的突变CAV毒株都能够产生一种中和性构象表位，这意味着它们可以引发所需的保护性免疫反应。这些特性使这些突变CAV毒株成为开发减毒CAV疫苗的潜在候选者。
