Speulman E, Bouchez D, Holub E B, Beynon J L
Department of Biological Sciences, Wye College, Ashford, Kent, UK.
Plant J. 1998 May;14(4):467-74. doi: 10.1046/j.1365-313x.1998.00138.x.
The disease resistance genes RPS2 of Arabidopsis and N of tobacco, among other recently cloned resistance genes, share several conserved sequences. Degenerate oligonucleotide primers, based on conserved sequences in the nucleotide binding site (NBS) and a weak hydrophobic domain of RPS2 and N, were used to amplify homologous sequences from Arabidopsis thaliana. Amplification products were obtained that were similar in sequence to the disease resistance genes RPS2, RPM1, N and L6. The Arabidopsis CIC-YAC library was used to identify the position of the disease resistance homologs on the Arabidopsis genome. Their map positions could be correlated with the disease resistance loci RPS5, RAC1, RPP9, CAR1, RPP7, RPW2, RPP1, RPP10, RPP14, RPP5, RPP4, RPS2, RPW6, HRT, RPS4, RPP8, RPP21, RPP22, RPP23, RPP24 and TTR1. This method was therefore not only successful in the identification of sequences located within gene clusters that are involved in disease resistance, but could also contribute to the cloning of disease resistance genes from Arabidopsis.
拟南芥的抗病基因RPS2和烟草的N基因，以及其他最近克隆的抗病基因，具有几个保守序列。基于RPS2和N基因核苷酸结合位点（NBS）中的保守序列以及一个弱疏水结构域设计的简并寡核苷酸引物，被用于从拟南芥中扩增同源序列。获得的扩增产物在序列上与抗病基因RPS2、RPM1、N和L6相似。利用拟南芥CIC-YAC文库确定抗病同源基因在拟南芥基因组上的位置。它们的图谱位置与抗病位点RPS5、RAC1、RPP9、CAR1、RPP7、RPW2、RPP1、RPP10、RPP14、RPP5、RPP4、RPS2、RPW6、HRT、RPS4、RPP8、RPP21、RPP22、RPP23、RPP24和TTR1相关。因此，该方法不仅成功鉴定了位于参与抗病的基因簇内的序列，还可能有助于从拟南芥中克隆抗病基因。
