A new method for rapid screening of bacterial species- or subspecies-specific DNA probes.
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.
Kook
Joong Ki
JK
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
Kim
Mi Kwang
MK
Seong
Jin Hyo
JH
Kim
Dong Kie
DK
Kim
Byung Ock
BO
Park
Joo Cheol
JC
Kim
Kack Kyun
KK
Choe
Son Jin
SJ
Min
Byung Moo
BM
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
D015342:DNA Probes; D004269:DNA, Bacterial; D004274:DNA, Recombinant; D004076:Digoxigenin