Yokoyama N, Maeda K, Kawaguchi Y, Ono M, Tohya Y, Mikami T
Department of Veterinary Microbiology, Faculty of Agriculture, University of Tokyo, Japan.
J Vet Med Sci. 1995 Aug;57(4):709-14. doi: 10.1292/jvms.57.709.
We constructed a recombinant virus containing defined deletion within thymidine kinase (TK) gene from feline herpesvirus type 1 (FHV-1) C7301, which was isolated in Japan, by standard rescue procedures with thymidine arabinoside (araT) selection. The araT resistant recombinant virus was designated as C7301dlTK. Southern blot analysis, together with polymerase chain reaction, revealed a deletion with the expected size, based on agarose gel electrophoresis, of the TK gene region in the C7301dlTK. Growth kinetics of the C7301dlTK in CRFK cells was similar to that of ther parent C7301 that possesses TK activity. However, plaques produced by the C7301dlTK were significantly smaller than those of the parent virus. This report migth be applicable for the development of a new genetically engineered FHV-1 vaccine.
我们通过使用阿糖胸苷(araT)选择的标准拯救程序,构建了一种重组病毒,该病毒包含来自1型猫疱疹病毒(FHV-1)C7301(在日本分离)胸苷激酶(TK)基因内的特定缺失。对araT具有抗性的重组病毒被命名为C7301dlTK。基于琼脂糖凝胶电泳,Southern印迹分析以及聚合酶链反应显示,C7301dlTK中TK基因区域存在预期大小的缺失。C7301dlTK在CRFK细胞中的生长动力学与具有TK活性的亲本C7301相似。然而,C7301dlTK产生的噬斑明显小于亲本病毒的噬斑。本报告可能适用于新型基因工程FHV-1疫苗的开发。