• 한국약용작물학회지
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• 제21권3호
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• pp.165-173
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• 2013

The fruits of Schisandra chinensis have been used as an edible ingredient and traditional medicine in Korea. Due to morphological similarities of dried mature fruits, the correct identification of S. chinensis from other closely related Schisandrae species is very difficult. Therefore, molecular biological tools based on genetic analysis are required to identify authentic Schisandrae Fructus. Random amplifed polymorphic DNA (RAPD) and Sequence Characterized Amplified Region (SCAR) were used to develop an easy, reliable and reproducible method for the authentication of these four species. In this paper, we developed several RAPD-derived species specific SCAR markers and established a multiplex-PCR condition suitable to discriminate each species. These genetic markers will be useful to distinguish and authenticate Schisandrae Fructus and four medicinal plants, S. chinensis, S. sphenanthera, S. repanda and K. japonica, in species level.

• Journal of Plant Biotechnology
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• 제50권
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• pp.34-44
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• 2023

멕시코 유래의 4배체 감자 근연야생종 중 하나인 Solanum hjertingii는 괴경에서 발생하는 흑변현상에 강한 것으로 알려져 감자의 신품종 육성에 유용한 형질로 이용이 가능하다. 이러한 저항성은 생리적 장해인 효소적 갈변과 흑반을 감소시킬 수 있다. 하지만, S. hjertingii와 S. tuberosum은 생리적 장벽에 기인한 교잡종 생산이 제한적인 관계로 직접적인 교배육종보다는 체세포잡종을 육성하는 방법을 활용할 수 있다. 체세포잡종 계통이 육성이 되면 분자표지를 이용한 적절한 잡종 계통을 선발하는 것이 필요하여, 본 연구에서는 S. hjertingii의 전체 엽록체 유전체 정보를 이용하여 S. hjertingii 특이적인 PCR 기반의 분자마커를 개발하였다. S. hjertingii의 전체 엽록체 유전체는 155,545 bp였으며, 다른 Solanum 종들과 구조 및 유전자 구성이 매우 유사하였고, 가지과의 다른 15개의 종들과 계통수 분석에서 근연야생종 S. demissum, S. hougasii, S. stoloniferum과 매우 가까운 유연관계를 나타냈다. 또한, S. hjertingii의 전체 엽록체 유전체와 8개의 다른 Solanum 종의 전체 엽록체 유전체의 다중 정렬 결과로 S. hjertingii 특이적인 1개의 InDel 영역과 7개의 SNP 영역을 확인하였고, 이를 이용하여 1개의 InDel 및 4개의 SNP 기반 PCR마커를 개발하였다. 본 연구의 결과는 S. hjertingii의 진화적 측면에서의 연구와 S. hjertingii를 이용한 감자의 신품종 육성 연구에 기여를 할 수 있을 것이다.

• 대한본초학회지
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• 제37권4호
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• pp.9-16
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• 2022

Objectives : In the Korean Pharmacopoeia 12th edition (KP 12) and the Korean Herbal Pharmacopoeia (KHP), two authentic herbal medicines are described, namely Bang-gi (Cheong-pung-deung) and Mok-bang-gi, respectively. In China, Bun-bang-gi is also used as herbal medicine. This study was conducted to develop a molecular authentication tool for distinguishing the three herbal medicine used as Bang-gi, which are Sinomeni Caulis et Rhizoma (Rhizome of Sinomenium acutum), Stephaniae Tetrandrae Radix (Root of Stephania terandra), and Cocculi Radix (Root of Cocculus trilobus). Methods : Twelve samples of three species (four samples of S. acutum, five samples of S. tetrandra, and three samples of C. trilobus) were collected from different habitats. The sequences of internal transcribed spacer (ITS) regions were obtained and comparatively analyzed to design the species-specific sequence characterized amplified region (SCAR) primers. The specificity of each pair of SCAR primers that amplified species-specific amplicon was evaluated for establishing the singleplex and multiplex PCR assay tools. Results : The singleplex SCAR markers show discriminability in C. acutum, S. tetrandra, and C. trilobus. These SCAR markers were also efficiently authenticated three species in the multiplex SCAR amplification using single PCR reaction. Furthermore, these PCR assay methods were applicable to authenticate dried herbal medicines distributed in the markets. Conclusions : The SCAR markers and PCR assay tools help discriminate the three herbal medicines used as Bang-gi at the species levels and provide a reliable genetic method to prevent the inauthentic distribution of these herbal medicines.

• 미생물학회지
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• 제39권1호
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• pp.36-39
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• 2003

본 연구에서는 16S rDNA복합중합효소연쇄반응을 이용하여 중이 삼출액에서 미생물병인원에 대한 특성을 알아보았다. 중이염환자의 중이 삼출액에서의 미생물 병인원은 주로 streptococcus pneumoniae, Haemophilus influenzae와 Moraxella catarrhalis이다. 26명의 환자로부터 39개의 중이염의 삼출액을 얻었고, 중이 삼출액에서 DNA를 추출하였다. PCR은 16S rDNA의 C4 region에서 21 base pair의 common primer와 각각 bacterium specific primers [(i) Haemophilus-specific primer (ii) Moraxella-specific primer and (iii) Streptococcus-specific primer]를 이용하여 수행하였다. 39 개의 중이염의 삼출액 시료 중에서, H. influenzae가 24 개(61.5%) 검출되었고, M. catarrhalis는 10 개(25.6%), S.pneumoniae는 3개 (7.7%)가 검출되었다. 16s rDNA 복합중합효소연쇄 반응 진단 결과,11 개(28%)의 중이삼출액 시료에서 음성을 나타내었다. 중이염의 중복감염은 9개의 중이 삼출액시료에서 관찰되었고, 이들은 모두 H.influenzae 와 M. catarrhalis 에 의한 중복감염이었다. 본 연구에서 저자 등은 165 rDNA 복합중합효소연쇄반응이 병원성 미생물을 빠르게 진단하고, 중이삼출액의 미생물 병인론을 추적할 수 있는 좋은 방법으로 제시하는 바이다.

• Asian-Australasian Journal of Animal Sciences
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• 제16권3호
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• pp.425-429
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• 2003

Staphylococcus aureus is a major pathogen for cattle, causing various forms of subclinical and clinical mastitis and could be a causative agent of food poisoning, it produces various superantigenic exotoxins which have a great public health significance. A total of 72 S. aureus clinical isolates from dairy farms located in Kyunggi Province Korea were examined for the species identification by biochemical method, and for the detection of $\beta$-lactamase, enterotoxin and other exotoxins genes by PCR. The results of species identification by biochemical method agreed with those of PCR done with species specific primer STA-AU. $\beta$-lactamase is an enzyme closely associated with the resistance to antibiotic penicillin, which is an important means of treatment of mastitis, all the isolates were positive for the presence of genes encoding $\beta$-lactamase, which were reproduced in penicillin susceptibility disc assay. Six types of toxin genes, Staphylococcal enterotoxin (SE)A, SEB, SEC, SEE, toxic shock syndrome toxin (TSST-1) and exfoliative toxin A (ET A) were detected in 72 isolates by PCR associated genotypic method in this study, none of the isolates carried the genes for enterotoxin D (SED) and exfoliative toxin B (ETB). The occurrence rate of exotoxin genes rated as 12.5%, and the precision of the PCR identification results has been confirmed using the reference strains.

• Fisheries and Aquatic Sciences
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• 제11권3호
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• pp.133-139
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• 2008

Molecular techniques, including restriction fragment length polymorphism(RFLP) and polymerase chain reaction-single strand conformational polymorph isms(PCR-SSCP), were developed to identify salted, dried threadfin(Eleutheronema tetradactylum) and white herring(Ilisha elongata) fish. Using PCR with universal primers, conserved 367-bp fragments of the cytochrome b gene were amplified from fresh fish samples and sequenced. The sequences were then searched for specific restriction sites. The digestion of the PCR products with the endonucleases AvaI, FokI, MboII, and MspI generated RFLP, which was used to identify the commercial products. Similarly, the amplified PCR-SSCP products were developed and the products tested. Overall, similar patterns were found in the majority of the fresh and processed products. Based on the results, both RFLP and PCR-SSCP were useful in determining and validating the authenticity of the fish species used to prepare the commercial salted, dried products. A similar approach can be applied to other species.

• 한국동물위생학회지
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• 제37권4호
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• pp.247-252
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• 2014

The economic impact of swine mycoplasma infection is high. An accurate diagnosis is often difficult and time consuming. We report the development and validation of an effective multiplex polymerase chain reaction (PCR) assay that detects Mycoplasma (M.) hyopneumoniae and M. hyorhinis. The multi detection of M. hyopneumoniae and M. hyorhinis primer set were employed to detect mycoplasma species and typing of the species was performed on the basis of sequence analysis of the PCR product. The target nucleic acid fragments were specifically amplified by M. hyopneumoniae and M. hyorhinis PCR with 16S ribosomal DNA primers. Single and mixed Mycoplasma species DNA templates were used to evaluate the specificity of the multiplex assay. The corresponding specific DNA products were amplified for each pathogen. The multiplex PCR assay provides a novel tool for simultaneous detection and differentiation of M. hyopneumoniae and M. hyorhinis.

• 한국균학회지
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• 제41권1호
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• pp.52-55
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• 2013

Phoma glomerata는 식물 잎이나 열매에 병을 일으키는 식물병원균으로 알려져 있다. 국내에서는 아직 피해사례가 없기 때문에 P. glomerata는 국내의 식물검역균으로 관리되고 있다. 본 연구는 국내에 들어오는 목재나 과일에 P. glomerata를 검출할 수 있는 방법 개발코자 수행되었다. Phoma 균주들의 translation elongation factor 1 alpha 유전자 염기서열에 기초하여 P. glomerata 특이적 PCR 프라이머를 디자인 하였고 그 특이성을 검정하였다. PCR 수행 결과 P. glomerata에서만 170 bp 크기의 밴드가 증폭되었고, 다른 비교 균주에서는 밴드가 증폭되지 않았다. 검출 감도를 평가하기 위해 기존 PCR방법과 real time PCR 방법을 이용하여 실험한 결과 최소 10 pg과 1 pg까지 각각 검출할 수 있었다. 본 연구결과는 디자인된 PCR 프라이머가 P. glomerata를 특이적으로 검출하는데 유용할 것임을 보여준다.

• Journal of Microbiology and Biotechnology
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• 제16권11호
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• pp.1778-1783
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• 2006

Event-specific PCR detection methods are the primary trend in genetically modified (GM) plant detection owing to their high specificity based on the flanking sequence of the exogenous integrant. Therefore, this study describes a real-time PCR system for event-specific GM canola GT73, consisting of a set of primers, TaqMan probe, and single target standard plasmid. For the specific detection of GT73 canola, the 3'-integration junction sequence between the host plant DNA and the integrated specific border was targeted. To validate the proposed method, test samples of 0, 1, 3, 5, and 10% GT73 canola were quantified. The method was also assayed with 15 different plants, and no amplification signal was observed in a real-time PCR assay with any of the species tested, other than GT73 canola.

• Mycobiology
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• 제30권4호
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• pp.202-207
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• 2002

This study was carried out to develop specific primers for PCR detection of Phellinus linteus. Diverse genomes of 15 Phellinus spp. including five Phellinus linteus isolates were fingerprinted by Primer Universal rice primer(URP)1F. The URP-PCR pattern differentiated P. linteus isolates from other phellinus spp. A polymorphic band(2.8 kb), which is unique for P. linteus isolates, was isolated and sequenced. Twenty four-oligonucleotide primer pairs were designed based on information of DNA sequence. The primer set(PLSPF2/PLSPR1) amplified single band(2.2 kb) of expected size with genomic DNA from seven Phellinus linteus, but not with that of other Phellinus species tested. The primers could be used identically in both DNA samples from mycelium and fruit bodies. This specific primers could offer a useful tool for detecting and identifying P. linteus rapidly.