• Development and Reproduction
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• v.22 no.2
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• pp.193-198
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• 2018

The author undertook PCR-founded genetic platform to investigate the hierarchical dendrogram of Euclidean genetic distances of one razor clam population, particularly for Solen corneus, which was further associated with those of the other clam population, by engaging with the precisely designed oligonucleotide primer sets. Seven oligonucleotides primers were used producing a total of 639 counted bands in population A and 595 in population B, respectively, ranging in size of DNA fragments from larger than approximately 50 bp to less than 1,100 bp. Their primers generated 39 specific fragments (6.10%) in population A and 47 (7.90%) in population B, respectively Comparatively, individuals of one razor clam population were fairly related to that of the other clam population, as shown in the hierarchical dendrogram of Euclidean genetic distances. The analysis of genetic variation between razor clam populations could offer important statistics for fisheries and mariculture. Generally the results showed specific and/or conserved genetic loci between razor clam populations. Specific markers established by the author will be valuable for the genetic analysis, species protection and increase of razor clam individuals in coastal region of the Korean Peninsula.

• Korean Journal of Ecology and Environment
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• v.33 no.3 s.91
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• pp.206-212
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• 2000

Since some cyanobacterial isolates under selective culturing conditions are lacking of characteristic specialized cells or showing altered morphology, the morpho-taxonomic criteria are not accurate enough to discriminate between species. Instead of morphological parameters, a method based on the single or the combination of repetitive oligonucleotides in a single PCR, repetitive oligonucleotides-primed PCR (ROP-PCR), was applied to generate DNA profiles for members of the cyanobacterial genera Anabaena and Oscillatoria, both of which are responsible for causing poisonous blooms in various freshwater systems. ROP-PCR performed on 10 isolates of the cyanobacteria with ERIC and REP sequences from gram-negative bacteria, STRR1A and LTRR sequences derived from cyanobacterial genome, and eukaryotic repetitive sequences, led to the identification of distinct genotypes, and provided specific and repeatable DNA fingerprints for cyanobacterial isolates. Grouping analysis of cyanobacterial isolates showed a signifiant difference depending on the primer used in PCR.

• The Plant Pathology Journal
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• v.20 no.3
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• pp.200-205
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• 2004

The partial nucleotide sequences of the genomic dsRNA mycovirus infecting Pleurotus ostreatus isolates ASI2223 and Suhan were determined and compared with those of mycoviruses belonging to partitiviruses and totiviruses. Partial nucleotide sequences of the purified dsRNA from ASI2223 and Suhan showed RNA-dependent RNA polymerase sequences that are closely related to those of partitiviruses, including Fusarium poae virus 1, Fusarium solani virus, Rhizoctoniasolani virus, Discula destructiva virus 2, and Oyster mushroom isometric virus 2. Specific primers were designed for RT-PCR detection of dsRNA viruses from the P. ostreatus isolate ASI2223 and Suhan. Two virus specific primer sets were found to specifically detect each virus among six sets of designed oligonucleotide primers. Collectively, these results suggest that dsRNA mycoviruses from P. ostreatus isolates ASI2223 and Suhan belong to the family Partitiviridae, although, they are not the same virus species. Our results also suggest that these virus-specific primer sets can be employed for the specific detection of each viral sequence in infected tissues.

• Journal of Life Science
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• v.33 no.10
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• pp.808-819
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• 2023

This study was conducted to develop a selection marker for the identification of the Bacillus licheniformis K12 strain in microbial communities. The strain not only demonstrates good growth at moderate temperatures but also contains enzymes that catalyze the decomposition of various polymer materials, such as proteases, amylases, cellulases, lipases, and xylanases. To identify molecular markers appropriate for use in a microbial community, a search was conducted to identify variable gene regions that show considerable genetic mutations, such as recombinase, integration, and transposase sites, as well as phase-related genes. As a result, five areas were identified that have potential as selection markers. The candidate markers were two recombinase sites (BLK1 and BLK2), two integration sites (BLK3 and BLK4), and one phase-related site (BLK5). A PCR analysis performed with different Bacillus species (e.g., B. licheniformis, Bacillus velezensis, Bacillus subtilis, and Bacillus cereus) confirmed that PCR products appeared at specific locations in B. licheniformis: BLK1 in recombinase, BLK2 in recombinase family protein, and BLK3 and BLK4 as site-specific integrations. In addition, BLK1 and BLK3 were identified as good candidate markers via a PCR analysis performed on subspecies of standard B. licheniformis strains. Therefore, the findings suggest that BLK1 can be used as a selection marker for B. licheniformis species and subspecies in the microbiome.

• Journal of Microbiology and Biotechnology
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• v.20 no.1
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• pp.193-201
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• 2010

A specific and rapid real-time PCR assay for detecting Ralstonia solanacearum in horticultural soil and plant tissues was developed in this study. The specific primers RSF/RSR were designed based on the upstream region of the UDP-3-O-acyl-GlcNAc deacetylase gene from R. solanacearum, and a PCR product of 159 bp was amplified specifically from 28 strains of R. solanacearum, which represent all genetically diverse AluI types and all 6 biovars, but not from any other nontarget species. The detection limit of $10^2\;CFU/g$ tomato stem and horticultural soil was achieved in this real-time PCR assay. The high sensitivity and specificity observed with field samples as well as with artificially infected samples suggested that this method might be a useful tool for detection and quantification of R. solanacearum in precise forecast and diagnosis.

• The Korean Journal of Mycology
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• v.46 no.2
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• pp.186-192
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• 2018

Rapid and accurate detection methods for Colletotrichum coccodes, an anthracnose pathogen of pepper and tomato, were developed using PCR. A specific primer set, coccoTef-F/coccoTef-R, which was constructed by analyzing tef-$1{\alpha}$ genes from 13 species and 22 strains of Colletotrichum, could specifically detect C. coccodes at a level of 10 ng by conventional PCR method and at 10 pg by real-time PCR. The PCR-based methods were also capable of detecting C. coccodes in pepper and tomato seeds artificially infected with the pathogen. The developed PCR methods can be applied for rapid and accurate inspection of C. coccodes in the seeds intended for export or import.

• Korean Journal of Veterinary Research
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• v.38 no.2
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• pp.345-352
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• 1998

The diagnosis of brucellosis is currently based on serological and microbiological tests. However, the microbiological isolation and identification have several disadvantages such as time-consuming and laborious, and the serological methods have been reported to cross-react with antigens other than those from Brucella spp. To develop a sensitive and rapid diagnostic method for detection of Brucella species, the genus-specific primers were designed and synthesized from the sequence of gene encoding a 31kDa cell surface protein(BCSP) and a 36kDa outer membrane protein(OMPB) of B abortus. The amplified 711bp and 982bp DNA fragments were only visible in each species of Brucella by PCR method using the BCSP and OMPB primers, respectively. However, PCR product was not obtained with DNA from other Gram-negative bacteria. As little as 1pg of the B abortus genomic DNA could be detected by this PCR method. Using the PCR technique, semen samples from 185 bulls of Brucella-seronegative herds in Cheju island were examined for comparison of this PCR method with conventional methods in 1995. The semen samples from 5 bulls were positive by culture method and PCR, and one was positive and 5 were suspect by semen plasma agglutination test. However, the semen samples obtained from 177 bulls were negative by semen plasma agglutination, culture and PCR methods in 1996. The results of comparison tests suggested that PCR was a better test than agglutination test against semen of bulls. This study indicated that the PCR technique was a valuable for the diagnosis of bovine brucellosis, particulary in bull semens.

• The Korea Journal of Herbology
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• v.29 no.6
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• pp.35-43
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• 2014

Objectives : Official Arisaematis Rhizoma is described only three species, Arisaema amurnse, Arisaema erubescens, and Arisaema heterophyllum, in national Pharmacopoeia. However, other Arisaema species, Arisaema ringens, Arisaema takesimense and Arisaema serratum, also have been distributed as an inauthentic Arisaematis Rhizoma in the herbal market. To develop a reliable molecular authentication method for Arisaematis Rhizoma in species level, we analyzed DNA barcode regions using six Arisaema species. Methods : Thirty-eight samples of six Arisaema plants species (A. amurense, A. amurense f. serratum, A. heterophyllum, A. takesimense, and A. serratum) were collected from different habitate and nucleotide sequences of DNA barcode regions (rDNA-ITS, matK, and rbcL gene) were analyzed after PCR amplification. The species-specific sequences and phylogenetic relations were estimated using entire sequences of three DNA barcodes based on the analysis of ClastalW and UPGMA, respectively. Results : The comparative analysis of DNA barcode sequences were revealed inter-species specific nucleotides to distinguish the medicinal plant of Arisaema Rhizoma in species levels excluding between A. amurense and its subspecies (A. amurense f. serratum) and A. takesimense and A. serratum, respectively. However, we obtained sequence differences enough to discriminate authentic and inauthentic Arisaematis Rhizoma. Therefore, we suggest that these SNP type molecular genetic markers were an reliable method avaliable to identify official herbal medicines. Conclusions : These marker nucleotides could be useful to identify the official herbal medicines by providing definitive information that can identify original medicinal plant and distinguish from inauthentic adulterants and substitutes.

• Plant Biotechnology Reports
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• v.4 no.1
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• pp.1-7
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• 2010

Definitive identification of original plant species is important for standardizing herbal medicine. The herbal medicines Cynanchi Wilfordii Radix (Baekshuoh in Korean and Beishuwu in Chinese) and Polygoni Multiflori Radix (Hashuoh in Korean and Heshuwu in Chinese) are often misidentified in the Korean herbal market due to morphological similarities and similar names. Therefore, we developed a reliable molecular marker for the identification of Cynanchi Wilfordii Radix and Polygoni Multiflori Radix. We used random amplified polymorphic DNA (RAPD) analysis of three plant species, Polygoni multiflorum, Cynanchum wilfordii, and Cynanchum auriculatum, to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed six sequence characterized amplification region (SCAR) markers for distinguishing Polygoni Multiflori Radix and Cynanchi Wilfordii Radix. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. These SCAR markers can be used for efficient and rapid authentication of these closely related species, and will be useful for preventing the distribution of adulterants.

• Journal of Food Hygiene and Safety
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• v.33 no.4
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• pp.306-309
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• 2018

The present study was conducted to investigate the prevalence of Mycoplasma spp. in cows and pigs with pneumonic lung lesions in Gyeonggi province in 2017. One hundred ninety two and 257 lung tissues were collected from slaughtered cows and pigs with pneumonic lesions, respectively, and examined for the presence of Mycoplasma spp. by a genus specific PCR. Among the examined animals, 147 cows (76.5%) and 203 pigs (80.9%) were found to be infected with Mycoplasma spp.. The infected tissues were further examined to identify the specific species of Mycoplasma using species specific PCRs. The only identified species in cows was M. agalactiae which was detected from 16 cows (8.3%), whereas M. dispar, M. bovis, and M. bovirhinis were not detected. In pigs, M. hyopneumoniae was detected from 74 pigs (28.8%) and M. hyorhinis from 13 pigs (5.1%). M. hyosynoviae was not detected. Taken together, the current study indicates Mycoplasma spp. are commonly associated with lung infection in cows and pigs in Korea. Further studies are needed to evaluate the impact of mycoplasma infection on the development of lung diseases in farm animals.