Frischmuth T, Stanley J
Universität Stuttgart, Biologisches Institut, Lehrstuhl für Molekularbiologie und Virologie der Pflanzen, Germany.
J Gen Virol. 1998 May;79 ( Pt 5):1265-71. doi: 10.1099/0022-1317-79-5-1265.
Nicotiana benthamiana was transformed with three different constructs (pCRA1, pCRA2 and pJC1) containing the coat protein coding sequence of African cassava mosaic virus (ACMV). Transformed plants were inoculated with a coat protein deletion mutant of ACMV that induces mild systemic symptoms in control plants. Several inoculated plants of transgenic lines CRA1/3, CRA1/4, CRA2/1 and CRA2/2 developed severe systemic symptoms typical of ACMV. DNA analysis revealed that, in these plants, recombination had occurred between the mutant viral DNA and the integrated construct DNA, resulting in the production of recombinant virus progeny with 'wild-type' characteristics. No reversion of mutant to 'wild-type' virus was observed in pJC1-transformed plants. Recombinant virus from several transgenic plants was analysed by PCR and parts of DNA A of virus progeny were cloned. Sequence analysis revealed that only a few nucleotides were changed from the published sequence.
用含有非洲木薯花叶病毒（ACMV）外壳蛋白编码序列的三种不同构建体（pCRA1、pCRA2和pJC1）转化本氏烟草。用ACMV的一种外壳蛋白缺失突变体接种转化植株，该突变体在对照植株中诱导轻度系统症状。转基因系CRA1/3、CRA1/4、CRA2/1和CRA2/2的几株接种植株出现了典型的ACMV严重系统症状。DNA分析表明，在这些植株中，突变病毒DNA与整合的构建体DNA之间发生了重组，导致产生具有“野生型”特征的重组病毒后代。在pJC1转化植株中未观察到突变体回复为“野生型”病毒。通过PCR分析了来自几株转基因植株的重组病毒，并克隆了病毒后代的部分DNA A。序列分析表明，与已发表序列相比只有少数核苷酸发生了变化。
