Hooft van Huijsduijnen R A, van Loon L C, Bol J F
Department of Biochemistry, State University of Leiden, Wassenaarseweg 64, 2333 AL Leiden, and Department of Plant Physiology, Agricultural University, Arboretumlaan 4, 6703 BD Wageningen, The Netherlands.
EMBO J. 1986 Sep;5(9):2057-61. doi: 10.1002/j.1460-2075.1986.tb04466.x.
A cDNA library was constructed to 10-15 S poly(A) RNA from tobacco mosaic virus (MV)-infected Samsun NN tobacco.By differential colony hybridization of 1400 transformants,32 clones were obtained corresponding to TMV-inducible tobacco mRNAs. These clones were subdivided into six clusters on the basis of cross-hybridization of the inserts. By Northern blot hybridization it was shown that three of the corresponding mRNAs were strongly induced by spraying tobacco plants with salicylic acid, whereas one mRNA was weakly induced by this treatment. All mRNAs were systemically induced in plants in which only the lower leaves were locally infected by TMV. Hybrid-selected translation was performed, using six clones representing one cluster each, followed by immunoprecipitation using an antiserum to purified pathogenesis-related (PR) proteins. Four clones yielded precipitable translation products. One of these clones represented a cluster of PR-1 clones, another clone encoded the thaumatin-like (TL) protein of tobacco which may correspond to PR-P or -Q.
构建了一个烟草花叶病毒(TMV)感染的三生NN烟草10 - 15S聚腺苷酸(poly(A))RNA的cDNA文库。通过对1400个转化子进行差异菌落杂交,获得了32个与TMV诱导的烟草mRNA相对应的克隆。根据插入片段的交叉杂交,这些克隆被细分为六个簇。通过Northern印迹杂交表明,用水杨酸喷洒烟草植株可强烈诱导其中三个相应的mRNA,而该处理对另一个mRNA的诱导较弱。在仅下部叶片被TMV局部感染的植株中,所有mRNA均被系统诱导。使用分别代表一个簇的六个克隆进行杂交选择翻译,然后用针对纯化的病程相关(PR)蛋白的抗血清进行免疫沉淀。四个克隆产生了可沉淀的翻译产物。其中一个克隆代表PR - 1克隆簇,另一个克隆编码烟草的类甜蛋白(TL),其可能对应于PR - P或 - Q。