• Applied Biological Chemistry
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• v.48 no.1
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• pp.77-81
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• 2005

To monitor GM soybean in soybean processed foods, tofu and biji, we prepared tofu and biji containing 0%, 1%, 3%, 5% and 100% GM soybean, respectively. We examined epsps gene inserted in soybean by PCR and EPSPS protein expressed in soybean using western blotting and lateral flow strip test to compare the sensitivity of these methods. A PCR product of 123 bp inserted in GM soybean was detected in all tofu and biji containing 1%, 3%, 5% and 100% GM soybean with the exception of 0% samples; however, the size of 600 bp inserted in GM soybean was only detected in tofu containing 100% soybean and in biji containing 5% and 100% soybean. In the protein level, GM soybean product was only detected in tofu and biji containing 100% GM soybean by western blotting. In addition, only biji containing 100% GM soybean was detected by lateral flow strip test. We concluded that in order to detect GM soybean efficiently in processed food, the PCR method is more sensitive than immunological methods. With the PCR method, small size product with approximately 100 bp in PCR product is sensitive to detect GM soybean in processed foods.

• Journal of Environmental Science International
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• v.20 no.4
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• pp.551-554
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• 2011

The production of the sharp-toothed eel by commercial catch off waters of Korea is annually declined after 1978. This study was carried out to obtain the stock management of the sharp-toothed eel using the PCR-aided RFLP method. The mtDNA COI gene was amplified using species-specific primers and PCR product was observed to 700 bp. Amplified DNA fragments were treated with six kinds of restriction enzymes (BaeHI, EcoRI, PstI, Ksp22, HinfI and HaeIII). The treatment of HaeIII showed a distinct PCR product between Yeosu/Jinhae/Jeju/Goseoung and Jangheung/Haenam populations that were observed from 300 to 400 bp in reference to 100 bp molecular marker. However, DNA fragment within populations had an identical pattern. The phylogenetic homology is 82% between two populations inferred from RFLP PCR product pattern using NTsysPC ver. 2.1. The use of HaeIII plays an important role in discriminating populations. It is thought that adults after over-wintering in the southern part of Jeju migrate to the Yeosu, Jinhae and Goseoung regions to spawn instead of to southwestern waters. Individuals within populations showed a relatively active genetic mixing and migration regardless of geography. However, the genetic ancestor of Jangheung and Haenam populations is appeared to be more adjacent to China or Japan than Jeju.

• Journal of Life Science
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• v.8 no.6
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• pp.673-680
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• 1998

Adenosine deaminase gene from Nocardioides sp. J-326TK was cloned by polymerase chain reaction using primers (PI, PII and PIII) constructed from the highly conserved amino acid sequences among Escherichia coli, mouse and human. A PCR product of about 800bp, as expected from the sequence of E. coli adenosine deaminase gene, was obtained from Nocardioides sp. J-326TK chromosomal DNA double-digested with EcoRI and Pst I. DNA sequencing of the PCR product after cloning into pT7Blue T-vector shows 99.5% and 98.9% homologies in nucleotide and amino acid sequences, respectively, with the E. coli adenosine deaminase whereas 59.5% and 46.8% homologies with the human adenosine deaminase, indicating the evolutionarily relationship of these organisms.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.6
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• pp.948-955
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• 1997

Immunomagnetic separation of virus coupled with .reverse transcription-polymerase chain reaction (IMS-PCR) was performed with infectious hematopoietic necrosis virus (IHNV). A DNA fragment of expected size was synthesized in the RT-PCR with total RNA extracted from IHNV inoculated CHSE-214. In a SDS-PAGE analysis, a protein band of over 70kDa was detected from non-infected cells and cells inoculated with IHNV and infectious pancreatic necrosis virus (IPNV). This protein was detected in the Western blot analysis probably because of non-specific reaction to monoclonal antibody against IHNV nucleocapsid protein. In the immunomagnetic separation, magnetic beads coated with monoclonal antibody against the IHNV nucleocapsid protein was incubated with supernatant from IHNV inoculated CHSE-214 cells. During this process, the non-specifically reacting protein could be removed by washing the magnetic bead with PBS in the presence of an external magnetic field, and viral proteins were detected from the remaining, cleaned magnetic beads. It was necessary to extract viral RNA from the captured virus particles before RT-PCR, and no DNA product was detected when the captured virus was only heated 5 min at $95^{\circ}C$. A PCR-product of expected size was synthesized from IMS-PCR with magnetic beads double coated either by goat anti-mouse IgG antibody -monoclonal antibody or streptavidin - biotin conjugated monoclonal antibody.

• The Plant Pathology Journal
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• v.18 no.1
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• pp.12-17
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• 2002

For the molecular detection of Sweet potaio feathery mottle virus (SPFMV) from diseased sweet potato plants, reverse transcription and polymerase chain reaction (RT-PCR) was performed with the use of a set of virus-specific primers to amplify an 816 bp product. The viral coat protein gene was selected for the design of the primers. No PCR product was amplified when Turnip mosaic virus, Potato vims Y or Cucumber mosaic virus were used as template in RT-PCR with the SPFMV-specific primers. The lowest concentration of template viral RNA required for detection was 10 fg. The vim was rapidly detected from total nucleic acids of leaves and roots from the virus-infected sweet potato plants as well as from the purified viral RNA by the RT-PCR. Twenty-four sweet potato samples were selected and analyzed by RT-PCR and restriction fragment length polymorphism (RFLP). RFLP analysis of the PCR products showed three restriction patterns, which resulted in some point mutations suggesting the existence of quasi-species for the vims in the infected sweet potato plants.

• Korean Journal Plant Pathology
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• v.14 no.6
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• pp.589-593
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• 1998

DNA amplification by polymerase chain reaction (PCR) was used to specifically detect Plasmodiophora brassicae, causing clubroot of crucifers. On the basis of DNA sequence informations, an oligonucleotide primer set specific for the pathogen was designed form small subunit gene (18S-like) and internal transcribed spacer (ITS) region of ribosomal DNA. Primer ITS 5/PB-C produced an amplification product of approximately 520 bp in length with DNA from P. brassicae. However, no amplification product was produced with DNAs from several soil-borne fungi, Didymella bryoniae and Rhizopus stolonifer. Using these primers, the clubroot pathogen was readily detected from infected roots of crucifers, but not from healthy roots. Southern hybridization analysis further confirmed that the amplification product was originated from P. brassicae.

• The Korean Journal of Mycology
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• v.44 no.2
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• pp.103-107
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• 2016

To establish a rapid and accurate detection of Phytophthora pinifolia, which is a quarantine pathogenic fungus in Korea, a species-specific primer was developed based on the ras-related protein (Ypt1) gene. Species-specific primer based on the DNA sequences of Ypt1 gene amplified 193 bp polymerase chain reaction (PCR) product for P. pinifolia. The primer pair yielded the predicted PCR product size exactly in testing with target pathogen DNAs, but not from the other 10 species of Phytophthora and 14 species of other phytopathogenic fungi. The primer pair also showed only the species-specific amplification curve on realtime PCR on target pathogen DNA. The detection sensitivity of real time PCR using species-specific primer pair was 10 to 100 times higher than conventional PCR, with 1 to $10pg/{\mu}L$.

• The Korea Journal of Herbology
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• v.29 no.3
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• pp.19-26
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• 2014

Objectives : Aucklandiae Radix (Muxiang) one of important herbal medicines in oriental medicine, is defined as the dried root of Aucklandia lappa (Asteraceae). Owing to the similarities in the morphology and name, Inulae Radix (Tu-Muxiang) and Vladimiriae Radix (Chuan-Muxiang) as well as Aristolochiae Radix (Qing-Muxiang) originated from other medicinal plants are often used as substitutes and/or adulterants of Aucklandiae Radix. Therefore, a reliable authentication of these herbal medicines is necessarily for the public health and prevention of misuse. Methods : 32 samples of medicinal plants supplying Aucklandiae Radix, Inulae Radix, Vladimiriae Radix, and Aristolochiae Radix were collected in Korea and China. The ITS (Internal transcribed spacer) nucleotide sequences of samples were determined. The PCR primers to amply DNA marker of each herbal medicine were designed basing on the specific ITS regions showing differences in the sequences among medicinal plants. Results : Primer set Al R/IS F designed in this work amplified 220 bp PCR product only in samples of Aucklandiae Radix. In contrast, primer set Ih F/IS R, Vs R/IS F, and AcR F1/Ac R amplified 250 bp product, 356 bp prouct, and 516 bp product respectively to identify Inulae Radix, Vladimiriae Radix, and Aristolochiae Radix. Conclusions : The primers designed basing on the nucleotide sequences of ITS regions appearing differenced in the sequences among medicinal plants amplified the DNA markers for the identification of Aucklandiae Radix, Inulae Radix, Vladimiriae Radix, and Aristolochiae Radix. These herbal medicines were more efficiently identified by multiplex PCR method using all primers in a single PCR process.

• Korean Journal of Plant Resources
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• v.14 no.1
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• pp.65-70
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• 2001

This study was performed to select marker which can identify genetic variation between mother plant and in vitro cultured plantlets of strawberry by PCR using random primer. When 'Yeobong' DNA extracted was treated with proteinase-K and RNase-H, clear DNA bands were shown. The optimal condition for RAPD in strawberry was to use 50ng of template DNA, 10pmol of primer,37oC of annealing temperature, and 45 cycles of PCR. After establishing above PCR optimal condition, RAPD pattern was investigated by using UBC primers. PCR was performed, and 46 of 90 primers produced PCR product showing 158 total bands. GC content was compared between the primers forming bands and no bands. The GC content showing bands was average 67.4%, whereas primers showing no bands 58%.

• Research in Plant Disease
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• v.17 no.3
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• pp.376-380
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• 2011

The bacterial speck of tomato caused by Pseudomonas syringae pv. tomato leads to serious economic losses especially on fruits of susceptible genotype. Thus, Pseudomonas syringae pv. tomato is a plant quarantine bacterium in many countries including Korea. In this study, we developed specific PCR assays for detection of the bacterium from tomato seeds. A specific primer set is designed from the hrpZ gene for specific detection of Pseudomonas syringae pv. tomato. A 501 bp PCR product corresponding to hrpZ gene was amplified only form Pseudomonas syringae pv. tomato strains, but no PCR product was amplified from other tomato bacterial pathogens, such as Pseudomonas syringae pv. glycinea, P. syringae pv. maculicola, P. syringae pv. atropurpurea, P. syringae pv. morsprunorum, and from other P. syringae pathovar strains. The nested-PCR primer set corresponding to an internal fragment of the 501 bp sequence (hrpZ) gine was used to specific detection of Pseudomonas syringae pv. tomato in tomato seed. A 119 bp PCR product using nested PCR primer was highly specific and sensitive to detect low level of Pseudomonas syrigae pv. tomato in tomato seeds. We believe that the PCR assays developed in this study is very useful to detect Pseudomonas syringae pv. tomato from the tomato seeds.