Parrish C R, Carmichael L E
Virology. 1986 Jan 15;148(1):121-32. doi: 10.1016/0042-6822(86)90408-3.
A mutant of canine parvovirus (CPV) was derived after culture of a highly passaged isolate of CPV in the NLFK feline cell line. The virus (CPV-102/10) differed from the parental strain in its antigenic type when tested with a panel of neutralizing monoclonal antibodies, and in its relative ability to replicate in a dog cell line or in dogs. The mutant formed little replicative form DNA in the canine cells. Five single recombinants and two double recombinants between CPV-102/10 and a wild type CPV were constructed by digesting purified viral replicative form DNA with restriction enzymes, ligating the separated ends from the different strains, and examining the viruses isolated after transfecting the DNA into cell cultures. Analysis of the recombinant viruses showed that the mutation(s) determining both the antigenic and host range differences mapped between 64 and 73 map units in the genome, within the capsid protein gene. Sequencing the DNA from that region revealed differences of two adjacent amino acids, both resulting in nonconservative differences in the predicted amino acid sequences of the viruses. These results show that the ability of CPV to infect dogs or their cultured cells is determined, at least in part, by the conformation of the surface of the virus capsid.
犬细小病毒(CPV)的一个突变体是在NLFK猫细胞系中对CPV的一个高度传代分离株进行培养后获得的。当用一组中和性单克隆抗体进行检测时,该病毒(CPV - 102/10)在抗原类型上与其亲代毒株不同,并且在狗细胞系或狗体内的相对复制能力也不同。该突变体在犬细胞中几乎不形成复制型DNA。通过用限制性内切酶消化纯化的病毒复制型DNA、连接不同毒株分离的末端,并检测将DNA转染到细胞培养物后分离出的病毒,构建了CPV - 102/10与野生型CPV之间的五个单重组体和两个双重组体。对重组病毒的分析表明,决定抗原性和宿主范围差异的突变位于基因组中衣壳蛋白基因内64至73个图谱单位之间。对该区域的DNA进行测序发现两个相邻氨基酸存在差异,这两个差异均导致病毒预测氨基酸序列中的非保守差异。这些结果表明,CPV感染狗或其培养细胞的能力至少部分由病毒衣壳表面的构象决定。