A new method for rapid screening of bacterial species- or subspecies-specific DNA probes |
| |
Authors: | Kook Joong Ki Kim Mi Kwang Seong Jin Hyo Kim Dong Kie Kim Byung Ock Park Joo Cheol Kim Kack Kyun Choe Son Jin Min Byung Moo |
| |
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 |
| |
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. |
| |
Keywords: | Inverted dot blot hybridization DNA probe Screening method |
本文献已被 PubMed 等数据库收录! |
|