Holt C A, Beachy R N
Department of Biology, Washington University, St. Louis, Missouri 63130.
Virology. 1991 Mar;181(1):109-17. doi: 10.1016/0042-6822(91)90475-q.
A full-length cDNA clone of the U1 (common) strain of tobacco mosaic virus (TMV) was constructed, and highly infectious transcripts were produced in vitro using bacteriophage T7 RNA polymerase. Frameshift mutations designed to cause premature termination of translation were introduced into either the 30-kDa movement protein (MP) gene or the coat protein (CP) gene. The MP-frameshift mutant was unable to locally or systemically infect inoculated tobacco plants. However, inoculation of transgenic tobacco plants that expressed a wild-type TMV MP gene resulted in both local and systemic viral infection. The CP-frameshift mutant, although unable to move systemically in nontransformed tobacco, exhibited systemic movement in transgenic plants that expressed a wild-type TMV CP gene. Transgenic tobacco plants that expressed the appropriate wild-type TMV gene were thus able to complement, in trans, mutant viruses lacking a functional MP or CP gene.
构建了烟草花叶病毒(TMV)U1(普通)株系的全长cDNA克隆,并使用噬菌体T7 RNA聚合酶在体外产生了高感染性转录本。将旨在导致翻译提前终止的移码突变引入30 kDa运动蛋白(MP)基因或外壳蛋白(CP)基因。MP移码突变体无法在接种的烟草植株上进行局部或系统感染。然而,对接种表达野生型TMV MP基因的转基因烟草植株后,病毒实现了局部和系统感染。CP移码突变体虽然无法在未转化的烟草中进行系统移动,但在表达野生型TMV CP基因的转基因植物中表现出系统移动。因此,表达适当野生型TMV基因的转基因烟草植株能够反式互补缺乏功能性MP或CP基因的突变病毒。