Salánki K, Balázs E, Burgyán J
Agricultural Biotechnology Center, Hungary.
Virus Res. 1994 Sep;33(3):281-9. doi: 10.1016/0168-1702(94)90109-0.
The nucleotide sequence of a cDNA clone corresponding to the entire RNA 3 of a pepper isolate of tomato aspermy cucumovirus (P-TAV) was determined. The 2222 nucleotides (nt) long RNA 3 is dicistronic and codes a 3a (280 amino acids) and a coat protein (cp, 218 amino acids). The nucleotide sequence of P-TAV RNA 3 shows a high sequence identity (99%) with that of C-TAV RNA 3 and the amino acid sequences of the 3a and the coat proteins between these two TAV strains are also very similar. The transcription start site of the subgenomic RNA 4 was mapped to the G at the position of 1148 of RNA 3. A full-length cDNA copy of P-TAV RNA-3 was cloned downstream of the phage T7 promoter. Transcripts synthesized in vitro had no extra nucleotides at the 5' end, but there were nine non-viral nucleotides at the 3' terminus. The transcripts of TAV RNA 3 were infectious when coinoculated into test plants in the presence of purified RNA 1 and 2 of P-TAV or Trk7-CMV respectively. Northern blot analysis of RNA extracted from inoculated plants showed the expected genomic and subgenomic RNAs.
测定了与番茄不孕黄瓜病毒辣椒分离株（P-TAV）的整个RNA 3对应的cDNA克隆的核苷酸序列。长度为2222个核苷酸（nt）的RNA 3是双顺反子的，编码一个3a蛋白（280个氨基酸）和一个外壳蛋白（cp，218个氨基酸）。P-TAV RNA 3的核苷酸序列与C-TAV RNA 3的核苷酸序列具有高度的序列同一性（99%），并且这两种TAV毒株之间的3a蛋白和外壳蛋白的氨基酸序列也非常相似。亚基因组RNA 4的转录起始位点被定位到RNA 3第1148位的G处。P-TAV RNA-3的全长cDNA拷贝被克隆到噬菌体T7启动子的下游。体外合成的转录本在5'端没有额外的核苷酸，但在3'末端有9个非病毒核苷酸。当分别与P-TAV或Trk7-CMV的纯化RNA 1和2共同接种到试验植物中时，TAV RNA 3的转录本具有感染性。对接种植物中提取的RNA进行Northern印迹分析，显示出预期的基因组RNA和亚基因组RNA。
