• Asian-Australasian Journal of Animal Sciences
• /
• 제20권12호
• /
• pp.1887-1894
• /
• 2007

In this paper, a rapid and reliable gene-targeted species-specific polymerase chain reaction (PCR) technique based on a two-step process was established to identify bifidobacteria in dairy products. The first step was the PCR assay for genus Bifidobacterium with genus specific primers followed by the second step, which identified the species level with species-specific primer mixtures. Ten specific primer pairs, designed from nucleotide sequences of the 16-23S rRNA region, were developed for the Bifidobacterium species including B. angulatum, B. animalis, B. bifidum, B. breve, B. catenulatum, B. infantis, B. longum, B. minimum, B. subtile, and B. thermophilum. This technique was applied to the identification of Bifidobacterium species isolated from 6 probiotic products, and four different Bifidobacterium spp. (B. bifidum, B. longum, B. infantis, and B. breve) were identified. The findings indicated that the 16S-23S rDNA gene-targeted species-specific PCR technique is a simple and reliable method for identification of bifidobacteria in probiotic products. PCR combined with Denaturing Gradient Gel Electrophoresis (DGGE) for identification of the bifidobacteria was also evaluated and compared with the gene-targeted species-specific technique. Results indicated that for fermented milk products consistency was found for both species-specific PCR and PCR-DGGE in detecting species. However, in some lyophilized products, the bands corresponding to these species were not visualized in the DGGE profile but the specific PCR gave a positive result.

• 한국약용작물학회지
• /
• 제13권2호
• /
• pp.121-125
• /
• 2005

An analysis of RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) was performed with three Angelica species (A. gigas Nakai, A. sinensis (Olive.) Diels and A. acutiloba Kitag) in an effort to distinguish between members of these three species. Two arbitrary primers (OPC02, OPD11) out of80 primers tested, produced 17 species-specific fragments among the three species. Eight fragments were specific for A. sinensis, four fragments specific for A. gigas, five specific for A. acutiloba. When primers OPC02 and OPD11 were used in the polymerase chain reaction, RAPD-PCR fragments that were specific for each of the three species were generated simultaneously. Primer OPC02 produced eight species-specific fragments: four were specific for A. sinensis, one for A. gigas, and three for A. acutiloba. Primer OPD11 produced nine speciesspecific fragments: four for A. sinensis, three for A. gigas, and two for A. acutiloba. The RAPD-PCR markers that were generated with these two primers should rapidly identify members of the three Angelica species. The consistency of the identifications made with these species-specific RAPD-PCR markers was demonstrated by the observation that each respective marker was generated from three accessions of each species, all with different origins. We also performed the RAPD-PCR analysis with the dried Angelica root samples that randomly collected from marketed and from the OPC02 primer, obtained a A. gigasspecific band and the band were cloned and sequenced.

• 한국발생생물학회지:발생과생식
• /
• 제23권4호
• /
• pp.367-375
• /
• 2019

Pufferfish (Takifugu spp.) are economically important edible marine fish. Mistakes in pufferfish classification can lead to poisoning; therefore, accurate species identification is critical. In this study, we used the mtDNA cytochrome c oxidase subunit I gene (COI) to design specific primers for six Takifugu species among the 21 domestic or imported pufferfish species legally sold for consumption in Korea. We rapidly and simultaneously identified these pufferfish species using a highly efficient, multiplex polymerase chain reaction (PCR) system with the six species-specific primers. The results showed that species-specific multiplex PCR (multiplex species-specific polymerase chain reaction; MSS-PCR) either specifically amplified PCR products of a unique size or failed. MSS-PCR yielded amplification fragment lengths of 897 bp for Takifugu pardalis, 822 bp for T. porphyreus, 667 bp for T. niphobles, 454 bp for T. poecilonotus, 366 bp for T. rubripes, and 230 bp for T. xanthpterus using the species-specific primers and a control primer (ca. 1,200 bp). We visualized the results using agarose gel electrophoresis to obtain accurate contrasts of the six Takifugu species. MSS-PCR analysis is easily performed and provides identification results within 6 h. This technique is a powerful tool for the discrimination of Takifugu species and will help prevent falsified labeling, protect consumer rights, and reduce the risk of pufferfish poisoning..

• Fisheries and Aquatic Sciences
• /
• 제26권11호
• /
• pp.678-688
• /
• 2023

Flatfish are one of the largest families in the order Pleuronectiformes and are economically important edible marine fish species. However, they have similar morphological characteristics leading to challenges in classifying correctly, which may result in mislabeling and illegal sales, such as fraudulent labeling of processed food. Therefore, accurate identification is important to ensure the quality and safety of domestic markets in Korea. Species-specific primers were prepared from the mainly consumed eleven species of the order Pleuronectiformes. To rapidly identify the 11 flatfish species, a highly efficient, rapid, multiplex polymerase chain reaction (PCR) with species-specific primers was developed. Species-specific primer sets were designed for the mitochondrial DNA cytochrome c oxidase subunit I gene. Species-specific multiplex PCR (MSS-PCR) either specifically amplified a PCR product of a unique size or failed. This MSS-PCR analysis is easy to perform and yields reliable results in less time than the previous Sanger sequencing methods. This technique could be a powerful tool for the identification of the 11 species b the family Pleuronectidae and can contribute to the prevention of falsified labeling and protection of consumer rights.

• 한국미생물·생명공학회지
• /
• 제39권1호
• /
• pp.70-76
• /
• 2011

Weissella 속 유산균 검출의 차이를 이용한 한국산 및 중국산 김치 판별의 가능성 검토를 위하여 Weissella 속 9종 균주의 PCR 검출법을 개발하였다. 종(species) 수준에서의 Weissella 속 균주의 특이적 PCR 검출을 위한 primer는 recN 유전자의 염기서열을 이용하여 선정하였으며, 김치로부터 W. cibaria, W. confusa, W. koreensis, W. soli를 모두 검출하기 위해서는 20 ng template DNA가 필요한 것으로 나타났다. 한국산 김치시료로부터는 W. cibaria, W. confusa, W. koreensis가 높은 빈도로 검출되었지만, W. soli는 검출되지 않았다. 한편 중국산 김치시료로부터는 이들 4종의 Weissella 속 균주들이 모두 검출되었다. 본 연구자들이 개발한 W. soli 특이적 PCR 검출은 현시점에서 중국산 김치의 원산지 판별법으로 적용되기에는 한계점을 가지고 있지만, 미생물 군집의 차이를 이용한 새로운 과학적 검증법이 제시되어 그 가능성이 검토되었다는 점에서 의의를 가지고 있다.

• Journal of Animal Science and Technology
• /
• 제50권1호
• /
• pp.9-18
• /
• 2008

본 연구에서는 조류에서 종 특이적인 DNA 염기서열변이를 검증하기 위하여 닭, 꿩, 칠면조, 메추리의 immunoglobulin light chain constant domain 유전자를 클로닝하여 DNA염기서열을 분석하였다. 종간에 구별이 가능한 DNA 염기서열변이가 위의 유전자에서 관찰되었다. PCR을 이용하여 종을 구별하기 위하여 종 사이에 특이적인 DNA 염기서열 부위에 한 쌍의 종 특이적인 프라이머를 제작하였다. 또한 비교실험을 위하여 이미 알려진 cytochrome b와 tapasin 유전자에서도 두 쌍의 종 특이적 프라이머를 제작하였다. PCR결과 세 쌍의 프라이머 모두 종 특이적으로 DNA를 증폭하였다. Immunoglobulin 유전자의 염기서열 변이를 이용한 종 특이적인 PCR 방법은 조류의 유전자원 보존을 위한 이종간 카이메라 연구에 유용하게 이용될 수 있을 것이다.

• Animal cells and systems
• /
• 제9권1호
• /
• pp.41-46
• /
• 2005

This study was conducted to reveal tree species preference and inter-specific difference of foraging behavior among four canopy-dwelling birds at forest dominated by Quercus mongolica, Acer pseudosieboldian and Carpinus cordata at 1,000 meters above sea level during breeding season of birds from 1995 to 1997 in Mt. Jumbongsan. Breeding birds were about 25 species and dominant birds were Erithacus cyane, Parus ater and Parus palustris. A relatively high number of bush-nesters can be a characteristic of breeding bird community at study area. Three gleaners (Tits, P. varius, P. palustris and P. ater) selectively preferred the trees irrespective of dominant tree species, whereas bark foragers (Nuthatch, Sitta europaea) utilized the dominant trees. The four birds showed significant inter-specific difference in use of foraging location, but the three tits did not show significant inter-specific difference in use of foraging maneuver and trees. Closely related tits may coexist with each other by inter-specific different use of foraging location determined by foliage structure and leaf arrangement.

• 미생물학회지
• /
• 제41권2호
• /
• pp.117-124
• /
• 2005

Rotifer와 병든 넙치 자어로부터 분리된 2개의 균주는 표현형적인 특성 확인 결과 Vibrio ichthyoenteri로 확인이 되었다. V. ichthyoenteri를 검출하기 위래 고감도 PCR 방법 개발을 하기 위래 V. ichthyoenteri 16S-23S rRNA intergenic spacer region(ISR)을 분석하였고, V. ichthyoenteri중 특이적 primer를 개발하였다. V. ichthyoenteri 의 ISR를 분석한 결과 1개의 다형성 ISR type서열을 포함하고 있었다. ISR서열은 길이는 348bp이며 tRNA gene을 가지고 있지 않았다. 이 서열을 가지고 이미 알려진 다른 Vibrio 종의 ISR 서열과 mutiple alignment를 수행한 결과 여러 영역에서 높은 가변성을 나타내어 가변 부위를 표적으로 하여 V. ichthyoenteri를 검출하기 위한 종 특이적 primer를 제작하였다. 제작된 primer의 특이성을 확인하기 위해 Vibrio 표준균주 19종의 genomic DNA와 분리균주 18 group에 genomic DNA 그리고 V. ichthyoenteri와 가장 유사한 서열을 가지고 있다고 알려진 Vibrio 종의 genomic DNA를 가지고 시험하였다. 그 결과 본 연구에서 제작된 종 특이적 primer를가지고 PCR 반응을 하면 V. ichthyoenteri를 검출 할 수가 있다.

• International Journal of Oral Biology
• /
• 제38권2호
• /
• pp.43-49
• /
• 2013

The objective of this study was to develop PCR primers that are specific for Streptococcus sanguinis, Streptococcus parasanguinis, and Streptococcus gordonii. We designed the S. sanguinis-, S. parasanguinis-, and S. gordonii-specific primers, Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 respectively, based on the nucleotide sequences of the Ssa21, Spa17, and Sgo41 DNA probes that were screened using inverted dot blot hybridization (IDBH). The species-specificity of these primers was assessed against 43 strains of mitis group streptococci, including clinical strains of S. sanguinis, S. parasanguinis, and S. gordonii. The resulting PCR data revealed that species-specific amplicons had been obtained from all strains of the target species tested, and that none of these amplicons occurred in any other strains from other species. These results suggest that the Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 primers may be useful in detecting S. sanguinis, S. parasanguinis, and S. gordonii at the species level, respectively.

• The Plant Pathology Journal
• /
• 제31권3호
• /
• pp.212-218
• /
• 2015

In this study, we developed a species-specific PCR assay for rapid and accurate detection of three Xanthomonas species, X. axonopodis pv. poinsettiicola (XAP), X. hyacinthi (XH) and X. campestris pv. zantedeschiae (XCZ), based on their draft genome sequences. XAP, XH and XCZ genomes consist of single chromosomes that contain 5,221, 4,395 and 7,986 protein coding genes, respectively. Species-specific primers were designed from variable regions of the draft genome sequence data and assessed by a PCR-based detection method. These primers were also tested for specificity against 17 allied Xanthomonas species as well as against the host DNA and the microbial community of the host surface. Three primer sets were found to be very specific and no amplification product was obtained with the host DNA and the microbial community of the host surface. In addition, a detection limit of $1pg/{\mu}l$ per PCR reaction was detected when these primer sets were used to amplify corresponding bacterial DNAs. Therefore, these primer sets and the developed species-specific PCR assay represent a valuable, sensitive, and rapid diagnostic tool that can be used to detect three specific pathogens at early stages of infection and may help control diseases.