Rakowski A G, Symons R H
Department of Plant Science, Waite Institute, University of Adelaide, Glen Osmond, Australia.
Virology. 1994 Sep;203(2):328-35. doi: 10.1006/viro.1994.1491.
Infectivity of linear monomeric viroid RNA transcripts is known to be dependent on the site of cloning of the viroid and the viroid and vector sequences at the termini of the RNA transcripts. We report here the further characterization of cloning site and sequence requirements using 11 different monomeric cDNA constructs of citrus exocortis viroid; these differed from one another by the site of cloning and by nonviroid residues adjacent to the 5' and 3' termini of the viroid sequence in the RNA transcript. The infectivity of these transcripts was assayed by inoculation onto tomato seedlings; all infectious transcripts were wild type, indicating accurate processing. The basic requirement for infectivity appeared to be the ability of the RNA transcripts to form a short double-stranded region of viroid and vector sequences at the junction of the two termini. The site of cloning of the viroid RNA used for cloning was not important as cDNA clones prepared at seven sites provided infectious transcripts.
已知线性单体类病毒RNA转录本的感染性取决于类病毒的克隆位点以及RNA转录本末端的类病毒和载体序列。我们在此报告使用11种不同的柑橘裂皮类病毒单体cDNA构建体对克隆位点和序列要求的进一步表征;这些构建体在克隆位点以及RNA转录本中类病毒序列5'和3'末端相邻的非类病毒残基方面彼此不同。通过接种到番茄幼苗上来检测这些转录本的感染性;所有感染性转录本均为野生型,表明加工准确。感染性的基本要求似乎是RNA转录本在两个末端的连接处形成类病毒和载体序列的短双链区域的能力。用于克隆的类病毒RNA的克隆位点并不重要,因为在七个位点制备的cDNA克隆都提供了感染性转录本。