J Korean Acad Periodontol.  2002 Jun;32(2):269-279. 10.5051/jkape.2002.32.2.269.

Study on isolation of Prevotella nigrescens 9336-specific DNA probes using random cloning method

• Gang SW
• Kim SH
• Kim DK
• Seong JH
• Kim BO
• Han JJ
• Kook JK

• KMID: 2057039
• DOI: http://doi.org/10.5051/jkape.2002.32.2.269

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.

Keyword

Prevotella nigrescens 9336; DNA probe

MeSH Terms

Base Sequence
Blotting, Southern
Clone Cells*
Cloning, Organism*
DNA Probes*
DNA*
Gene Library
Mass Screening
Polymerase Chain Reaction
Prevotella nigrescens*
Prevotella*
DNA
DNA Probes