摘要
目的研究一种在微生物遗传背景不清楚情况下，快速制备探针的新方法。方法采用随机扩增ＤＮＡ多态性技术随机扩增李斯特菌基因组，将其中的单核细胞增生李斯特菌扩增产物克隆制备ＤＮＡ探针，探针用聚合酶链反应标记上３２Ｐ，与李斯特菌和非李斯特菌进行菌落原位杂交鉴定其特异性。结果获得一长度为８５０ｂｐ，能与所有李斯特菌特异杂交而不与非李斯特菌反应的李斯特菌属特异探针。结论随机扩增ＤＮＡ多态性技术为快速制备微生物ＤＮＡ探针提供了一种新途径。
Objective To develop a universal method to construct DNA probe specific for a given microorganism without any genetic background knowledge of the target species.Methods Chromosomal DNA of Listeria spp were amplified by RAPDPCR. The product from L. monocytogenes was cloned into PGEM TVector and the cloned probes were labeled with 32P by PCR and hybridized with genome DNA from different species of Listeriec and related bacteria other than Listeria.Results a 850bp DNA probe was obtained with the specificity only for Listeria spp. Conclusions RAPDPCR method is of valuable in constructing DNA probe specific for microorganism intelested, avoiding the tedious procedure of constructing and screening of genome libraries as is required with previously described means of isolating such probes.
