• Korean Journal of Microbiology
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• v.36 no.4
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• pp.273-278
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• 2000

The aacCl, aacC2, aacC3, and aacC4 genes, which encode aminoglycoside acetyltransferase AAC(3)-I, AA(3)-II, AAC(3)-III, and AAC(3)-IV, respectively, and aerolysin genes in water samples from Juam lake were surveyed by polymerase chain reaction. Surface water was collected from January 1996 to December 1998, and then bacterial DNA was extracted from the water. Twelve samples were tested by PCR to servey the genes for aminoglycoside acetyltransferase and aerolysin in the lake water. The aacC2 gene was detected in 9 of 12 DNA samples. Among 9 samples showing aacC2 positive, 7 samples were associated with Tn3 sequence. However, none of the twelve samples amplified the expected DNA fragment for aacC1, aacC3, and aacC4 genes. PCR primer to detect the aerolysin gene was designed using the conserved region of the genes for aerolysin and hemolysin of Aeromonas spp. This primer set successfully amplified the expected 414 bp PCR product with the DNA samples from the lake water. The aerolysin gene was detected in 7 of 12 DNA samples. When Southern hybridization of the gel with probe was performed, the aerolysin gene was detected in 10 of 12 DNA samples. However, the seasonal fluctuation of these genes was not found.

• Korean Journal of Animal Reproduction
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• v.20 no.3
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• pp.323-331
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• 1996

Predetermination of sex in mammalian species has many aspects of application including the prenatal diagnoses of genetic disorders in humans and sex-selected breeding programs in the animal industry. Embryos sexing can be carried out using the polymerase chain reaction (PCR) to amplify specific sequences present in the sex chromosomes, or by fluorescent in situ hybridization (FISH) of specific probes to the X and Y chromosomes. A 3.3 kb porcine male-specific DNA fragment (pEM39) was cloned previously in our laboratory. In this study, FISH and PCR methods were employed to examine if the pEM39 can be used a sex-specific DNA probes Porcine ovaries were obtained from a local slaughter house and oocytes collected. All oocytes were subjected to in vitro maturation followed by 1n vitro fertilization. Parthenogenetically activated embryos were served as a negative control. Embryonic samples were collected at the 2-cell stages and PCR was performed to analyze DNA. Among 10 embryos examined, four embryos were identified as males and six were females. The cloned male-specific DNA fragment showed male-specificity for the cells in the liver tissue and the porcine early embryos by FISH. It was also demonstrated that the cloned male-specific DNA is localized on the hetero chromatic region of the long arm in the Y chrom-osome (Yq) as shown by the FISH and karyotyping. The results suggest that the cloned male-specific DNA fragment may be useful for predetermination of sex with a few embryonic cells. The porcine male-specific sequence can be a reliable index for embryo sexing by PCR.

• Journal of Plant Biology
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• v.35 no.3
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• pp.253-257
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• 1992

Potyvirus group is the largest group among plant virus groups and damages severely plant hosts upon infectiQn. In order to investigate the mechanism by which potyviruses induce disease in plants, a cDNA clone 29-6 which is cOIlsidered to be a cDNA clone for garlic mosaic virus (GMV) was isolated. It did not hybridize to garlic latent virus genome, which is one of two major garlic viruses. Northern blot analysis shows that the genome size of garlic mosaic virus was about 9 kb. Clone 29-6 strongly hybridizes to poly(A) RNA isolated from garlic leaves, suggesting that GMV RNA is polyadenylated as other potyviruses. Nucleotide sequence analysis of cDNA clones overlapping with clone 29-6 showed that garlic plants are infected with various strains of garlic mosaic virus which are closely related to each other. other.

• Bulletin of the Korean Chemical Society
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• v.31 no.11
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• pp.3099-3102
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• 2010

A label-free DNA detection method was developed for a simple electrochemical DNA sensor with a short assay time. Self-assembled monolayers of peptide nucleic acid were used as a probe on gold electrodes. The formation of the self-assembled monolayers on the gold electrodes was successfully checked by means of cyclic voltammetry. The target DNA, hybridized with peptide nucleic acid, can be detected by the anodic peak current of ferrocene dendrimers, which interact electrostatically with the target DNA. This anodic peak current was measured by square wave voltammetry at 0.3 V to decrease the detection limit on the order of the nanomolar concentrations. As a result, the label-free electrochemical DNA sensor can detect the target DNA in concentrations ranging from 1 nM to $1\;{\mu}M$ with a detection limit of 1 nM.

• Journal of Korean Society of Forest Science
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• v.83 no.1
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• pp.20-24
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• 1994

In woody species with a long life span, the studies on inheritance of any trait may be very time consuming and laborious. Chloroplast DNA(cpDNA) has been a valuable tool in such studies since it has several unique features such as limited genome size and cytoplasmic inheritance. In the present study, cpDNAs from five different species of Populus(P. alba, P. glandulosa, P. alba${\times}$P. glandulosa, P. davidiana, and P. nigra), and Nicotiana tabacum were compared with regard to restriction fragment length polymophism. The results showed that cpDNAs among the species were very conserved, although some polymorphisms were observed when the DNAs were digested with restriction enzyme EcoRI or KphI. The other enzymes (Bgl II, and PstI) tested produced identical restriction fragmentation pattern among the species. However, cpDNAs from all the five Populus species showed different restriction fragmentation pattern from that of tobacco with the four restriction enzymes tested. Southern hybridization with tobacco rbcL gene fragment as a probe also produced identical pattern among Populus species. The results indicate that cpDNAs in the genus are very well conserved during evolution.

• Applied Biological Chemistry
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• v.36 no.6
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• pp.547-552
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• 1993

Soybean nuclear extracts and S-100 were prepared to examine the soybean embryo factors which bind to the upstream region of soybean ${\beta}-conglycinin$ ${\alpha}'$ subunit gene. SEF3(soybean embryo factor 3), which is presumed to be a trans-acting factor for the expression of the gene, was detected in gel mobility shift assay using the DNA probe containing two AACCCA hexanucleotides. DNA probe containing CATGCAT or AACACA was used to find any other soybean embryo factor interacting with the upstream region of ${\beta}-Conglycinin$ ${\alpha}'$ subunit gene. It was found that there was no common DNA binding protein detected both in nuclear extracts and S-100. The relative levels of SEF3 binding activity both in nuclear extracts and S-100 of maturing soybean seeds were determined. SEF3 activity of nuclear extracts was first detected around 20 days after pollination and significantly increased around 32 days after pollination.

• KSBB Journal
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• v.21 no.5
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• pp.347-352
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• 2006

This study was carried out to establish the optimum condition for cell disruption with a sonificator in the detection of the gram positive bacteria, Bacillus globigii and Streptococcus epidermidis for the purpose of developing automatic fluorometer. The efficiency of sonificator on the Bacillus globigii and Streptococcus epidermidis disruption differed greatly according to the diameter of sonificator probe tip. The larger sonificator probe diameter showed greater disruption. Bacillus globigii was more disruptive than Streptococcus epidermidis. Sonificator probe of the 13 mm diameter was the most efficient one when sample was sonificated for 20 seconds. The detection limits of Bacillus globigii and Streptococcus epidermidis were $10^5CFU/m{\ell}\;and\;5{\times}10^5CFU/m{\ell}$ respectively when samples were sonificated for 20 seconds with a sonificator probe of 13 mm diameter.

• Fisheries and Aquatic Sciences
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• v.12 no.3
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• pp.227-235
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• 2009

The marine toxic dinoflagellate Alexandrium catenella has been implicated in numerous paralytic shellfish poisoning (PSP) events in many countries. Due to difficulties in rapidly identifying A. catenella, field-based study of this species has been problematic. The present study developed a TaqMan format A. catenella-specific probe for real-time PCR assay (specific to Korean genotype) based on LSU rDNA sequence information for studying geographic and temporal distribution of the species in surface sediments and water columns of Jinhae Bay, Korea. The field survey from 2007 to 2008 revealed that A. catenella occurred in most seasons at low densities, mostly below 1 cell $mL^{-1}$, and was more abundant in spring (maximum cell density of 2 cells $mL^{-1}$) when shellfish exceed the quarantine toxin level for PSP toxins in Jinhae Bay.

• Bulletin of the Korean Chemical Society
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• v.29 no.5
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• pp.943-947
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• 2008

A phosphate-specific and fluorescent probe was prepared for label-free phosphatase assays based on fluorescence polarization. By using the probe, dephosphorylation reactions of DNA and protein substrates by calf intestinal alkaline phosphatase (CIP) could effectively be monitored in real-time. Since this assay method does not require additional materials such as labeled substrates and phosphospecific antibodies to obtain fluorescence polarization signals, it is simple, cost-effective, and expected to be useful not only for measuring activity of phosphatases but also for high-throughput screening of phosphatase inhibitors.

• Mycobiology
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• v.50 no.5
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• pp.382-388
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• 2022

White mold (or Sclerotinia stem rot), caused by Sclerotinia species, is a major air, soil, or seed-transmitted disease affecting numerous crops and wild plants. Microscopic or culture-based methods currently available for their detection and identification are time-consuming, laborious, and often erroneous. Therefore, we developed a multiplex quantitative PCR (qPCR) assay for the discrimination, detection, and quantification of DNA collected from each of the three economically relevant Sclerotinia species, namely, S. sclerotiorum, S. minor, and S. nivalis. TaqMan primer/probe combinations specific for each Sclerotinia species were designed based on the gene sequences encoding aspartyl protease. High specificity and sensitivity of each probe were confirmed for sclerotium and soil samples, as well as pure cultures, using simplex and multiplex qPCRs. This multiplex assay could be helpful in detecting and quantifying specific species of Sclerotinia, and therefore, may be valuable for disease diagnosis, forecasting, and management.