Infectivity of recombinant strawberry vein banding virus DNA
Infectivity of recombinant strawberry vein banding virus DNA
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DOI：
10.1099/vir.0.18994-0
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发表时间：
2003-06-01
影响因子：
3.8
通讯作者：
Mahmoudpour, A
中科院分区：
文献类型：
--
作者：
Mahmoudpour, A
利用基因枪技术对草莓镶脉病毒（SVBV）基因组进行了克隆，并以4周龄草莓（Fragaria vesca L.）变种UC-5）植物的细胞中，金颗粒包被有推定的全长7-9 kb病毒DNA。接种后6-7周，15%的接种植株出现脉带症状。大约1（.）25-将病毒DNA的一个片段克隆到双元载体pCGN 1547中。将该构建体的粒子轰击到草莓植物中得到75%的感染率。将该构建体用于根癌农杆菌的转化，并且将这些细胞渗入健康的草莓叶中导致在100%的接种植物中出现脉带症状。凝胶电泳，Southern印迹杂交与SVBV探针和PCR扩增的DNA片段的序列分析被用来确认SVBV感染症状的植物。
Infectivity of the cloned DNA genome of strawberry vein banding virus (SVBV) was demonstrated by particle bombardment of 4-week-old strawberry (Fragaria vesca L. var. UC-5) plants with gold particles coated with the putative full-length 7-9 kb viral DNA. Vein banding symptoms developed on 15 % of inoculated plants 6-7 weeks post-inoculation. An approximate 1(.)25-mer of the viral DNA was cloned into the binary vector pCGN 1547. Particle bombardment of this construct into strawberry plants gave an infection rate of 75 %. The construct was used for transformation of Agrobacterium tumefaciens, and infiltration of these cells into healthy strawberry leaves resulted in development of vein banding symptoms in 100 % of inoculated plants. Gel electrophoresis, Southern blot hybridization with an SVBV probe and sequence analyses of PCR-amplified DNA fragments were used to confirm SVBV infection in symptomatic plants.