Friedrich Miescher Institut, P.O. Box 2543, CH-4002, Basel, Switzerland.
Plant Mol Biol. 1986 Sep;6(5):303-12. doi: 10.1007/BF00034937.
A hybrid Cauliflower Mosaic Virus (CaMV) genome containing a selectable marker gene was constructed by replacing the gene VI coding region with the aminoglycoside (neomycin) phosphotransferase type II [APH(3')II] gene from Tn5. This modified viral genome was tested for its infectivity both in planta and in a protoplast transformation system of Brassica campestris var. rapa. Stable, genetically transformed cell lines of B. campestris var. rapa were obtained after transformation. DNA of the hybrid CaMV genome was found to be integrated into high molecular weight plant genomic DNA. Transformation was achieved only when the hybrid genome was supplied together with wild type viral DNA. A possible complementation of the modified CaMV genome with the wild type viral DNA as a helper molecule in planta and in the protoplast system is discussed.
构建了一种含有选择标记基因的杂种 Cauliflower Mosaic Virus（CaMV）基因组，方法是用来自 Tn5 的氨基糖苷（新霉素）磷酸转移酶 II [APH(3')II]基因取代基因 VI 编码区。该修饰后的病毒基因组在拟南芥植物体内和原生质体转化系统中进行了感染性测试。成功获得了稳定的、经遗传转化的油菜变种甘蓝型油菜细胞系。杂种 CaMV 基因组的 DNA 被发现整合到高分子量的植物基因组 DNA 中。只有当提供杂种基因组和野生型病毒 DNA 时，才能实现转化。讨论了修饰后的 CaMV 基因组与野生型病毒 DNA 作为辅助分子在植物体内和原生质体系统中的可能互补作用。
