• Proceedings of the Plant Resources Society of Korea Conference
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• 2002.04a
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• pp.33-33
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• 2002

• Korean Journal of Plant Resources
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• v.16 no.1
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• pp.55-60
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• 2003

The genetic variation of random amplified polymorphic DNA (RAPD) patterns of genomic DNAs was investigated in dioecious plant Rumex acetosa L., which carries different sex chromosome complements in female (2n=12A+XX) and male (2n=12A+XY$_1$Y$_2$). One hundred and twenty random primers consisted of 10-mer were used for PCR amplification. Polymorphic bands were found in 24 primers. Specific bands for female and male were 16 and 18, respectively. Especially, a band of 1,440 bp from the OPC-10 primer was male specific. These sex specific RAPD markers are used to understanding the sex determination mechanism in plants.

• Fisheries and Aquatic Sciences
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• v.6 no.1
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• pp.13-19
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• 2003

Genomic DNA from the muscle of marsh clam (Corbicula leana) from Gochang, Muan and a Chinese site was extracted to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction (RAPD- PCR). Out of 20 primers, seven primers produced amplified fragments which were consistently polymorphic. A total of 1,246 amplified products were produced of which 530 were polymorphic $(42.5\%)$. The number of polymorphic bands produced per primer varied from 40 to 122 with an average of 75.7 in marsh clam from Gochang. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic. Also, about $4.34\%$ of total polymorphic bands were specific to marsh clam from Gochang. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, which were polymorphic. This common bands in every individuals should be diagnostic of specific strains, species and/or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of 12 specific bands. The intra-population variation was revealed in the band patterns identified by this primer. The random primer OPB-12 (CCTTGACGCA) yielded the amplified fragments which were consistently polymorphic between the marsh clams from Gochang and from Muan. This primer produced a total of 77 polymorphic bands: 31 bands from Gochang, 14 from Muan and 32 from the Chinese populations. An average of polymorphic bands were 1.8 from Gochang and 2.5 from the Chinese populations. This value obtained from the Chinese population was higher than those from the two domestic populations. Generally, the RAPD polymorphism generated by these primers may be useful as a genetic marker for strain or population identification of marsh clam.

• The Plant Pathology Journal
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• v.32 no.1
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• pp.53-57
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• 2016

Major diseases in grafted cacti have been reported and Fusarium oxysporum, Bipolaris cactivora, Phytophthora spp. and Collectotrichum spp. are known as causal pathogens. These pathogens can lead to plant death after infection. Therefore, some European countries have quarantined imported cacti that are infected with specific fungal pathogens. Consequently, we developed PCR detection methods to identify four quarantined fungal pathogens and reduce export rejection rates of Korean grafted cacti. The pathogen specific primer sets F.oF-F.oR, B.CF-B.CR, P.nF-P.nR, and P.cF-P.CR were tested for F. oxysporum, B.cactivora, P. nicotinae, and P. cactorum, respectively. The F.oF-F.oR primer set was designed from the Fusarium ITS region; the B.CF-B.CR and P.nF-P.nR primers respectively from Bipolaris and Phytophthora ITS1; and the P.cF-P.CR primer set from the Ypt1protein gene region. The quarantine fungal pathogen primer pairs were amplified to the specific number of base pairs in each of the following fungal pathogens: 210-bp (F. oxysporum), 510-bp (B. cactivora), 313-bp (P. nicotinae), and 447-bp (P. cactorum). The detection limit for the mono- and multiplex PCR primer sets was 0.1 ng of template DNA under in vitro conditions. Therefore, each primer set successfully diagnosed contamination of quarantine pathogens in export grafted cacti. Consequently, our methodology is a viable tool to screen contamination of the fungal pathogen in exported grafted cacti.

• The Korean Journal of Malacology
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• v.32 no.4
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• pp.255-261
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• 2016

The selected seven oligonucleotides primers BION-32, BION-33, BION-35, BION-38, BION-40, BION-46 and BION-58 generated the shared loci, specific loci, unique shared loci to each population and shared loci by the three T. granosa populations in Beolgyo, a Chinese site and Wonsan, respectively. The bandsharing value between individuals' no. 03 and no. 04 was 0.816, which was the highest value identified within the Beolgyo population. The primer BION-35 generated the most loci (a total of 70), with an average of 10.0 in the Wonsan population. On average, seven oligonucleotides primers generated 16.1 specific loci in the Beolgyo population, 22.3 in the Chinese population and 39.3 in the Wonsan population. 126 unique shared loci to each population, with an average of 18 per primer, were observed in the Beolgyo population, 63 loci, with an average of 9 per primer, were observed in the Chinese population, and 49 loci, with an average of 7 per primer, and were observed in the Wonsan population. The oligonucleotides primer BION-32 generated 14 unique loci to each population, which were identifying each population in the Beolgyo population. Interestingly, every primer had not distinguished the shared loci by the three populations, major and/or minor fragments of sizes, which were identical in almost all of the samples. As regards average bandsharing value (BS) results, individuals from Beolgyo population ($0.717{\pm}0.057$) exhibited higher BS values than did those from Wonsan population ($0.552{\pm}0.104$) (P < 0.05). The dendrogram resulted from truthful seven oligonucleotides primers, representing three genetic clusters comprising group I (BEOLGYO 01, 02, 03, 04, 05, 06 and 07), group II (CHINESE 08, 09, 10, 11, 12, 13 and 14) and group III (WONSAN 15, 16, 17, 18, 19, 20 and 21). In three T. granosa populations, the longest genetic distance (0.874) displaying significant molecular difference was also between individual no. 02 within the Beolgyo population and individual no. 12 within the Chinese population. Relatively, individuals of the CHINESE population were fairly closely related to those of the WONSAN population.

• Journal of Applied Biological Chemistry
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• v.52 no.1
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• pp.1-7
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• 2009

For the development of qualitative PCR detection method of genetically modified (CM) rice, rice species-specific gene, OsCc-1 (rice cytochrome c gene), was selected as suitable far use as an endogenous gene in rice. The primer pair OsCytC-5'/3'with 111 bp amplicon was used for PCR amplification of the rice endogenous gene, OsCc-1 and no amplified product was observed from 8 different crops as templates. Qualitative PCR method was carried out with stack traits of L528$\times$CryIAc1 GM rice developed in Korea. For the qualitative PCRs, some primer pairs were designed with a construct-specific and event-specific type based on T-DNA and junction sequences of T-DNA in GM rice. Actck-5'/3' amplifying between actin promoter and OsCK1 gene introduced in LS28 gave rise to an amplicon 306 bp; also, CrLB-5'/3' from CryIAcl and CKRB-5'/3'amplifying the junction region of T-DNA and genome sequence from LS28 as event-specific primers gave rise to an amplicon 142 bp and 91 bp, respectively. These primer pairs for the detection of event-specific targets not produced PCR amplicons on non-CM rice and various crops in contrast to event lines. Therefore, in this study we verified that event-specific primers were effective to specifically detect stack trait lines and demonstrated that this method presented could be provided with the detection-method data for risk assessment analysis of GM rice to be commercialized.

• The Plant Pathology Journal
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• v.27 no.4
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• pp.367-371
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• 2011

Multiplex PCR assays were developed for the simultaneous detection of ten important Korean quarantine phytoplasmas. The species-specific primers were designed based on ribosomal protein, putative preprotein translocase Y, immunodominant protein, elongation factor TU, chaperonin protein and the 16S rRNA genes of 'Candidatus (Ca.) Phytoplasma' species. Three main primer sets were prepared from ten designed primer pairs to limit nonspecific amplification as much as possible. The multiplex PCR assay using the three primer sets successfully amplified the correct conserved genes for each 'Ca. Phytoplasma' species. In addition, ten important 'Ca. Phytoplasma' species could be easily determined by recognizing band patterns specific for each phytoplasma species from three primer sets. Moreover, a high sensitivity of multiplex PCR for each primer set was observed for samples containing a low DNA concentration (10 ng/${\mu}l$). This study provides the useful multiplex PCR assay as a convenient method to detect the presence of ten important quarantine phytoplasmas in Korea.

• Biomedical Science Letters
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• v.24 no.3
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• pp.230-238
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• 2018

Human enteric Adenovirus-41 (HuEAdV-41) causes gastroenteritis, which detected by the polymerase chain reaction (PCR) base diagnostic system for clinical, food, environmental, fish and shellfish samples. We developed improved PCR and nested PCR primer set which had high specificity, sensitivity and reduced times. In this study, we compared seventeen conditions reported in the previous study that was using the PCR based HuEAdV-41 detection system, and non-enteric Adenovirus were detected in nine conditions. The most sensitive detection condition was up to 25 copies however it took 184 minutes of PCR reaction time. In this study, the PCR primer set developed had same level of sensitivity, it reduced the time of detection for clinical, food and seafood samples to 112 minutes. Developed nested PCR primer set needed 112 minutes but detected up to approximately 1 copy. In addition, developed PCR and nested PCR primer set was validated with twenty samples of underground water at random, of which ten samples showed specific band without non-specific reaction. We expect this study will be used to diagnose HuEAdV-41 from various samples.

• Journal of Plant Biotechnology
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• v.31 no.1
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• pp.1-6
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• 2004

Bang-Poong and related species are an important herbal medicine. However, it is difficult to determine the commercial dry material through anatomical and chemotaxonomical characteristics. Here, we used a PCR-based technique for an accurate discrimination of Bang-Poong and related species. With the RAPD primers, 215 RAPDSs(random amplified polymorphic DNAs) were obtained, and 98% of them showed polymorphic patterns. RAPDs from the four primers were appropriate for the discrimination of S. divaricata $(T_{URCZ{\cdot}})\;S_{CHISKIN}$, those from the six primers for P. japonicum $T_{HUNBERG}$, those from the four primers for P. terebinthaceum $F_{ISHER}$, and those from the six primers for G. littoralis Fr. $S_{CHMIDT}$. The specific bands from the primer 425 were obtained and used to develop SCAR (sequence characterized amplified region) markers, based on the sequence information of the RAPD markers. The SCAR primers generated a 215 bp fragment specific to Peucedanum terebinthaceum $F_{ISHER}$, and a 177 bp and a 300 bp fragment specific to G. littoralis Fr. $S_{CHMIDT}$. As a result, the three SCAR markers were able to discriminate from two Bang-Poong related species.

• Korean Journal of Microbiology
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• v.48 no.4
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• pp.309-313
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• 2012

The purpose of this study is to simultaneously detect and identify specific genus of Lactobacillus distributed in vagina of healthy women in their twenties by using multiplex primer. Vaginal fluid samples were taken from 166 women who were healthy and did not have vaginosis symptoms caused by bacterial infection. Six species of lactic acid bacteria belong to Lactobacillus genus that are known to inhabit in healthy vagina were selected to make a species-specific multiplex primer. Multiplex primer I was specified to selectively detect L. iners, L. crispatus, L. gasseri and multiplex primer II was specified to selectively detect L. acidophilus, L. jensenii, L. fermentum. L. crispatus (77%) was most frequently detected, and L. acidophilus (57%) and L. jensenii (57%) were relatively higher than others. Although the proportion of L. iners (59%) was a little higher than those of L. acidophilus and L. jensenii, this should be further validated in healthy women's vagina to be sure since they often appear in women having bacterial vaginosis and/or during the antibiotic therapy. Conclusively, the multiplex PCR technique using the species-specific primer could a useful tool to predict variation of vaginal health condition and process of recovery from vaginal disease.