Saarela M, Stucki A M, von Troil-Lindén B, Alaluusua S, Jousimies-Somer H, Asikainen S
Institute of Dentistry, University of Helsinki, Finland.
FEMS Immunol Med Microbiol. 1993 Mar;6(2-3):99-102. doi: 10.1111/j.1574-695X.1993.tb00310.x.
Genetic analysis of 31 clinical strains of Porphyromonas gingivalis isolated from nine subjects, 2-6 strains per subject, was performed by Southern hybridization. Chromosomal DNA was extracted by the method of Moncla et al. [1] and digested to completion with restriction endonucleases PstI, ClaI and BglI. The DNA fragments were separated electrophoretically on agarose gels, transferred to nylon membranes and hybridized to the non-radioactively labelled plasmid pKK 3535 which contains the rmB ribosomal RNA operon of the Escherichia coli chromosome. Of the three enzymes, BglI was the most suitable for the genetic analysis of P. gingivalis. With this enzyme, the intra-individual strains were shown to be identical in eight of the nine subjects, whereas inter-individual strains were different.
通过Southern杂交对从9名受试者中分离出的31株牙龈卟啉单胞菌临床菌株(每名受试者2 - 6株)进行了基因分析。染色体DNA采用Moncla等人[1]的方法提取,并用限制性内切酶PstI、ClaI和BglI完全消化。DNA片段在琼脂糖凝胶上进行电泳分离,转移到尼龙膜上,然后与含有大肠杆菌染色体rmB核糖体RNA操纵子的非放射性标记质粒pKK 3535杂交。在这三种酶中,BglI最适合用于牙龈卟啉单胞菌的基因分析。使用这种酶时,9名受试者中有8名受试者的个体内菌株显示相同,而个体间菌株则不同。