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KMID : 0363020020320020001
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Journal of Korean Academy of Periodontology
2002 Volume.32 No. 2 p.1 ~ p.21
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Study on isolation of Prevotella nigrescens 9336 - specific DNA probes using random cloning method
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강순원
김세훈/김동기/성진효/김병옥/국중기
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Abstract
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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.
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KEYWORD
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Prevotella nigrescens 9336, DNA probe
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