Linthorst H J, Danhash N, Brederode F T, Van Kan J A, De Wit P J, Bol J F
Department of Biochemistry, Leiden University, The Netherlands.
Mol Plant Microbe Interact. 1991 Nov-Dec;4(6):586-92. doi: 10.1094/mpmi-4-586.
Clones corresponding to tobacco pathogenesis-related (PR) proteins PR-4 and tomato PR protein P2 were isolated from phage cDNA libraries of tobacco infected with tobacco mosaic virus and tomato infected with Cladosporium fulvum, respectively. The probe used in these screenings was a polymerase chain reaction product, synthesized on phage DNA from the tobacco cDNA library, using a synthetic oligonucleotide primer whose sequence corresponded to the partial amino acid sequence available for P2. The different cDNA sequences from the tobacco and tomato clones contained open reading frames for small proteins with 80-90% amino acid sequence identity. Both tobacco PR-4 and tomato P2 are synthesized as precursor proteins, with an N-terminal signal peptide involved in extracellular targeting. The proteins are highly similar to putative wound-induced proteins of potato (win) and to the precursor protein of hevein. However, in contrast to the hevein pro-protein and win proteins, PR-4 and P2 do not contain N-terminal, chitin-binding "hevein" domains. The tobacco and tomato genomes contain a limited number of genes corresponding to PR-4 or P2, whose expression is induced upon infection with the above-mentioned pathogens.
分别从感染烟草花叶病毒的烟草噬菌体cDNA文库和感染番茄叶霉病菌的番茄噬菌体cDNA文库中分离出了与烟草病程相关蛋白PR-4以及番茄PR蛋白P2相对应的克隆。这些筛选中使用的探针是一种聚合酶链反应产物,它是利用一个合成寡核苷酸引物在来自烟草cDNA文库的噬菌体DNA上合成的,该引物的序列与P2可用的部分氨基酸序列相对应。来自烟草和番茄克隆的不同cDNA序列含有小蛋白的开放阅读框,其氨基酸序列同一性为80-90%。烟草PR-4和番茄P2均以前体蛋白形式合成,带有一个参与细胞外靶向的N端信号肽。这些蛋白与马铃薯假定的创伤诱导蛋白(win)以及橡胶素前体蛋白高度相似。然而,与橡胶素原蛋白和win蛋白不同的是,PR-4和P2不包含N端几丁质结合“橡胶素”结构域。烟草和番茄基因组中与PR-4或P2相对应的基因数量有限,其表达在感染上述病原体后被诱导。