title: A new method for rapid screening of bacterial species- or subspecies-specific DNA probes.
abstract: A simple assay for the rapid screening of bacterial species- or subspecies-specific DNA probes for the random cloning method is presented, involving the use of genomic DNAs as probes and recombinant plasmid DNAs containing genomic DNA digested with HindIII as targets. The optimal amount of target DNAs and the concentration of digoxigenin-labeled genomic DNA probes were 20 ng and 100 ng ml(-1) (or 10 ng and 200 ng ml(-1)), respectively. The method was applied to the development of Fusobacterium nucleatum subspecies-specific probes. Our results showed that four out of 96 probes were F. nucleatum subspecies-specific, which was confirmed by Southern blot analysis. Our results indicate that the new method can be used for the rapid screening of species- or subspecies-specific probes.
authors.affiliation: Department of Oral Biochemistry, College of Dentistry, Chosun University, 375 Seo-Suk Dong, Dong-ku, 501-759, Gwang-ju, South Korea. jkkook@chosun.ac.kr
authors.forename: Joong Ki
authors.initials: JK
authors.lastname: Kook
authors.forename: Mi Kwang
authors.initials: MK
authors.lastname: Kim
authors.forename: Jin Hyo
authors.initials: JH
authors.lastname: Seong
authors.forename: Dong Kie
authors.initials: DK
authors.lastname: Kim
authors.forename: Byung Ock
authors.initials: BO
authors.lastname: Kim
authors.forename: Joo Cheol
authors.initials: JC
authors.lastname: Park
authors.forename: Kack Kyun
authors.initials: KK
authors.lastname: Kim
authors.forename: Son Jin
authors.initials: SJ
authors.lastname: Choe
authors.forename: Byung Moo
authors.initials: BM
authors.lastname: Min
mesh_terms: D015373:Bacterial Typing Techniques; D003001:Cloning, Molecular; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis; D004274:DNA, Recombinant / Q000032:analysis; D004076:Digoxigenin / Q000378:metabolism; D016680:Genome, Bacterial; D006090:Gram-Negative Bacteria / Q000145:classification* / Q000235:genetics; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010957:Plasmids; D013045:Species Specificity
chemical_list: D015342:DNA Probes; D004269:DNA, Bacterial; D004274:DNA, Recombinant; D004076:Digoxigenin