Study on isolation of Prevotella nigrescens 9336-specific DNA probes using random cloning method.
10.5051/jkape.2002.32.2.269
- Author:
Soon Won GANG
1
;
Se Hoon KIM
;
Dong Kie KIM
;
Jin Hyo SEONG
;
Byung Ock KIM
;
Jin Ju HAN
;
Joong Ki KOOK
Author Information
1. Department of Periodontology, College of Dentistry, Chosun University, Korea.
- Publication Type:Original Article
- Keywords:
Prevotella nigrescens 9336;
DNA probe
- MeSH:
Base Sequence;
Blotting, Southern;
Clone Cells*;
Cloning, Organism*;
DNA Probes*;
DNA*;
Gene Library;
Mass Screening;
Polymerase Chain Reaction;
Prevotella nigrescens*;
Prevotella*
- From:The Journal of the Korean Academy of Periodontology
2002;32(2):269-279
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
The purpose of this study is to develop species-specific DNA probes and polymerase chain reaction (PCR) primers for detection and identification of Prevotella nigrescens (P. nigrescens) 9336. This study procedure includes (1) whole-genomic DNA extraction of P. nigrescens 9336 (2) construction of the genomic DNA library, (3) screening of strain-specific DNA probe by reverse dot Hybridization method, (4) confirmation of strain-specific DNA probe by Southern blot analysis, (5) determination of nucleotide sequences of strain-specific DNA probe. Thirty-five restriction fragments of P. nigrescens 9336 genomic DNA digested with the Hind III were obtained. Reverse dot hybridization and Southern blot analysis data showed that three of them, Pn10, Pn23, and Pn35, could be P. nigrescens 9336-specific DNA probes. These data indicated that these DNA probes could be useful in detection and identification of the P. nigrescens 9336.
