Matzeit V, Schaefer S, Kammann M, Schalk H J, Schell J, Gronenborn B
Max-Planck-Institut für Züchtungsforschung, Abt. Genetische Grundlagen der Pflanzenzüchtung, Köln, Federal Republic of Germany.
Plant Cell. 1991 Mar;3(3):247-58. doi: 10.1105/tpc.3.3.247.
Wheat dwarf virus (WDV) is a geminivirus that infects monocotyledonous plants. To exploit the potential of WDV as a replicative gene vector, we developed a transient replication and expression system based on the transfection of protoplasts derived from Triticum monococcum suspension culture cells. Cloned genomic copies of various WDV isolates as well as mutants constructed in vitro were introduced into the protoplasts and assayed for their ability to replicate. As a result, regions of the WDV genome necessary or dispensable for the viral DNA replication could be defined. In addition, the gene encoding the viral capsid protein was replaced by three different bacterial marker genes, neomycin phosphotransferase, chloramphenicol acetyltransferase, and beta-galactosidase. The beta-galactosidase gene doubled the size of the WDV genome. The replication of the recombinant WDV genomes and the expression of these genes were monitored in suspension culture cells of T. monococcum. The potential of replicative expression vectors based on the WDV genome is discussed.
小麦矮缩病毒（WDV）是一种感染单子叶植物的双生病毒。为了开发WDV作为复制型基因载体的潜力，我们基于对来自一粒小麦悬浮培养细胞的原生质体进行转染，开发了一种瞬时复制和表达系统。将各种WDV分离株的克隆基因组拷贝以及体外构建的突变体导入原生质体，并检测它们的复制能力。结果，可以确定WDV基因组中对于病毒DNA复制必需或非必需的区域。此外，病毒衣壳蛋白编码基因被三个不同的细菌标记基因，即新霉素磷酸转移酶、氯霉素乙酰转移酶和β-半乳糖苷酶所取代。β-半乳糖苷酶基因使WDV基因组大小增加了一倍。在一粒小麦的悬浮培养细胞中监测重组WDV基因组的复制以及这些基因的表达。讨论了基于WDV基因组的复制型表达载体的潜力。
