Scott D L, Clark C W, Fyffe A E, Walker M D, Deah K L
US Department of Agriculture, Agriculture Research Service, Beltsville, MD 20705, USA.
Lett Appl Microbiol. 1998 Jul;27(1):39-44. doi: 10.1046/j.1472-765x.1998.00382.x.
The goal of this study was to develop a polymerase chain reaction (PCR) capable of differentiating Phytophthora species that are pathogenic on potatoes using a single primer pair. To achieve this objective, primers were derived from conserved regions flanking variable sequences in the internal transcribed spacer 1 (ITS1) of Phytophthora species. One primer pair produced a 140 bp product from P. infestans, P. erythroseptica and P. nicotianae. The PCR products were purified and used in an asymmetric PCR (A-PCR) protocol to generate single-strand DNA (ssDNA). The ssDNA of the Phytophthora potato pathogens reproducibly migrated in non-denaturing polyacrylamide gels in a species-specific manner.
本研究的目标是开发一种聚合酶链反应（PCR），该反应能够使用单一引物对区分对马铃薯致病的疫霉属物种。为实现这一目标，引物来源于疫霉属物种内部转录间隔区1（ITS1）可变序列侧翼的保守区域。一对引物从致病疫霉、粉红疫霉和烟草疫霉中扩增出一条140 bp的产物。PCR产物经纯化后用于不对称PCR（A-PCR）方案以生成单链DNA（ssDNA）。马铃薯疫霉病原体的ssDNA在非变性聚丙烯酰胺凝胶中以物种特异性方式可重复迁移。
