Merits Andres, Fedorkin Oleg N, Guo Deyin, Kalinina Natalia O, Morozov Sergey Yu
Institute of Biotechnology, Viikki Biocenter, University of Helsinki, PO Box 56, FIN-00014 Helsinki, Finland2.
National Institute of Chemical Physics and Biophysics, Akadeemia tee 23, EE12618 Tallinn, Estonia1.
J Gen Virol. 2000 Dec;81(Pt 12):3099-3106. doi: 10.1099/0022-1317-81-12-3099.
The putative replication initiation protein (Rep) of Coconut foliar decay virus (CFDV) was expressed as a 6x His recombinant protein in E. coli and in recombinant baculovirus. Purified 6x His-Rep protein was demonstrated to possess sequence non-specific RNA- and ssDNA-binding activities as well as magnesium-dependent ATPase/GTPase activity. The yeast two-hybrid system revealed that CFDV Rep could interact with itself. Subcellular distribution of the CFDV Rep was studied by fractionation of insect cells infected with recombinant baculovirus expressing the 6x His-Rep protein and by laser scanning confocal microscopy of Nicotiana benthamiana epidermal cells bombarded with a construct encoding CFDV Rep fused to GFP. It was shown that CFDV Rep associated predominantly with nuclei and membranes of infected/transfected cells. These activities of CFDV-encoded Rep are very similar to those reported for Reps of geminiviruses.
椰子叶枯病毒（CFDV）的假定复制起始蛋白（Rep）在大肠杆菌和重组杆状病毒中作为6x His重组蛋白表达。纯化的6x His-Rep蛋白被证明具有序列非特异性RNA和单链DNA结合活性以及镁依赖性ATPase/GTPase活性。酵母双杂交系统显示CFDV Rep可与自身相互作用。通过对感染表达6x His-Rep蛋白的重组杆状病毒的昆虫细胞进行分级分离，以及对用编码与GFP融合的CFDV Rep的构建体轰击的本氏烟草表皮细胞进行激光扫描共聚焦显微镜观察，研究了CFDV Rep的亚细胞分布。结果表明，CFDV Rep主要与感染/转染细胞的细胞核和膜相关。CFDV编码的Rep的这些活性与双生病毒Reps报道的活性非常相似。
