• Journal of Microbiology and Biotechnology
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• v.12 no.3
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• pp.508-513
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• 2002

Human endogenous retroviral long terminal repeats (LTRs) have been found to be coexpressed with sequences of genes located nearby. It has been suggested that the LTR elements have contributed to the structural change or genetic variation of human genome connected to various diseases. The HERV-W family has been identified in the cerebrospinal fluids and brains of individuals with schizophrenia. Using a cDNA library derived from a human brain, the HERV-W LTR elements were examined and five new LTR elements were identified. These elements were examined using a YAC clone panel from the Xq21.3 region linked to psychosis that was replicated on the Y chromosome after the separation of the chimpanzee and human lineages. Fourteen elements of the HERV-W LTR were identified in that region. Those LTR elements showed a high degree of sequence similarity ($91.8-99.5\%$) with previously reported HERV-W LTR. A phylogenetic tree obtained from the neighbor-joining method revealed that new HERV-W LTR elements were closely related to the AXt000960, AF072504, and AF072506 from the GenBank database. The data indicates that several copy numbers of the HERV-W LTR elements exist on the Xq21.3 region and are also expressed in the human brain. These LTR elements need to be further investigated as potential leads to neuropsychiatric diseases.

• BMB Reports
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• v.39 no.5
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• pp.578-585
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• 2006

The cDNA of a firefly luciferase from lantern mRNA of Lampyroidea maculata has been cloned, sequenced and functionally expressed. The cDNA has an open reading frame of 1647 bp and codes for a 548-residue-long polypeptide. Noteworthy, sequence comparison as well as homology modeling showed the highest degree of similarity with H. unmunsana and L. mingrelica luciferases, suggesting a close phylogenetic relationship despite the geographical distance separation. The deduced amino acid sequence of the luciferase gene of firefly L. maculata showed 93% identity to H. unmunsana. Superposition of the three-dimensional model of L. maculata luciferase (generated by homology modeling) and three dimensional structure of Photinus pyralis luciferase revealed that the spatial arrangements of Luciferin and ATP-binding residues are very similar. Putative signature of AMP-binding domain among the various firefly species and Lampyroidea maculata was compared and a striking similarity was found. Different motifs and sites have been identified in Lampyroidea maculata by sequence analysis. Expression and purification of luciferase from Lampyroidea maculata was carried out using Ni-NTA Sepharose. Bioluminescence emission spectrum was similar to Photinus pyralis luciferase.

• The Plant Pathology Journal
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• v.32 no.2
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• pp.102-111
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• 2016

Olive culture is very important in the Mediterranean Basin. A severe outbreak of Olive Quick Decline Syndrome (OQDS) caused by Xylella fastidiosa infection was first noticed in 2013 on olive trees in the southern part of Apulia region (Lecce province, southern Italy). Studies were carried out for detection and diversity evaluation of the Apulian strain of Xylella fastidiosa. The presence of the pathogen in olive samples was detected by PCR amplifying the 16S rDNA, gyrase B subunit (gyrB) and HL hypothetical protein genes and single nucleotide polymorphisms (SNPs) assessment was performed to genotype X. fastidiosa. Twelve SNPs were recorded over gyrB and six SNPs were found for HL gene. Less variations were detected on 16S rDNA gene. Only gyrB and HL provided sufficient information for dividing the Apulian X. fastidiosa olive strains into subspecies. Using HL nucleotide sequences was possible to separate X. fastidiosa into subspecies pauca and fastidiosa. Whereas, nucleotide variation present on gyrB gene allowed separation of X. fastidiosa subsp. pauca from the other subspecies multiplex and fastidiosa. The X. fastidiosa strain from Apulia region was included into the subspecies pauca based on three genes phylogenetic analyses.

• Journal of the Korean Society of Fisheries and Ocean Technology
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• v.59 no.3
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• pp.253-262
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• 2023

To identify sand crab Ovalipes punctatus populations and establish management units for each population, mtDNA COI regions were analyzed. As a result, the clade of O. punctatus in Korea were separated by two with a genetic distance of 0.17-2.08%, and there was no significant difference in the result of pairwise FST values representing genetic differentiation by sampling areas (p > 0.05). Also, no geographical separation found in the distribution of haplotypes and the results of the haplotype network. This result suggests that O. punctatus larvae were dispersed for a long time by the ocean current by suffering meroplanktonic period for 1 month, and increased the gene flow due to the development of the swimming legs for the increase in mobility. Therefore, in the results of mtDNA COI region analysis of O. punctatus in the East Sea, Yellow Sea, South Sea and East China Sea (Ieodo) of Korea, no clear intra-species differentiation was found.

• Journal of Applied Biological Chemistry
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• v.44 no.2
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• pp.92-94
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• 2001

A simple protocol for the detection of opines, cucumopines and mikimopines using a general horizontal or vertical get electrophoresis system for protein or DNA separation in the laboratory are demonstrated. This electrophoresis method can also be applied to other opines as long as correct detection reagent and buffer system are used.

• KSBB Journal
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• v.18 no.3
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• pp.190-196
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• 2003

An analytical system detecting DNA particularly utilizing a concept of membrane strip chromatography initially applied to home-version tests for, such as, pregnancy and ovulation has been developed. We have chosen S. typhimurium as model analyte among food-contaminating microorganisms that occurred in high frequencies, and invA gene, as a detection target, specific to Salmonella species. This gene was able to be amplified by PCR under optimal conditions employing newly designed primers in our laboratory. The PCR product was specifically measured via hybridization between the analyte and a DNA probe, which was a totally different feature from the conventional gel electrophoresis detecting the products based only on the molecular size. It is notable thar the DNA probe sequence was specially designed such that no separation of excess primers present after PCR was required. This was immobilized on a nitrocellulose (NC) membrane via streptavidin-biotin linkage minimizing a steric effect when the hybridization with the amplified DNA took place. The analyrical system detected the microorganism in a concentration of minimum $10^3$ cfu/mL (i.e., 10 cells per system), estimated from the standard curve, 20 to 40 minutes after adding the sample. This sneitivity was approximately 10 times higher than that of gel electrophoresis as an analytical tool conventionally used. Furthermore, the assay was able to be run at room temperature, which would ofter an extra advantage to users.

• Journal of the Korean Chemical Society
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• v.48 no.5
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• pp.483-490
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• 2004

We have developed a novel polymerase chain reaction (PCR)-microchip gel electrophoresis (MGE) method based on the sieving gel mixture of commercially available poly(vinylpyrrolidone) (PVP) and hydroxy ethyl cellulose (HEC) for the rapid detection and diagnosis of the orf virus (ORFV) from Korean indigenous goat. After amplification of 594-bp DNA fragment from the B2L gene of ORF virus, the amplicon was analyzed by the MGE separation. The glass microfluidic chip (64 mm total length (36 mm effective length)${\times}$90 ${\mu}$m width${\times}$20 ${\mu}$m depth) allowed the fast detection and diagnosis of ORFV in the mixture of 1.0% PVP ($M_r$ 360,000) and 1.0% HEC ($M_r$250,000) as a sieving matrix with better resolution and reproducibility of DNA fragments. Under the electric field of 277.8 V/cm, the 594-bp DNA was analyzed within 4 min. Compared to traditional slab gel electrophoresis, the PCR-MGE method was twenty times faster and an effective clinical method for the quantitative analysis of ORFV.

• Korean Journal of Microbiology
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• v.44 no.1
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• pp.22-28
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• 2008

Cutting oils are emulsionable fluids widely used in metal working processes. Their composition is mineral oil, water, and additives (fatty acids, surfactants, biocides, etc.) generating a toxic waste after a long use. Cutting oils also affect colour, taste and odour of water, making it undesirable for domestic and industrial uses. In these days, conventional treatment methods as evaporation, membrane separation or chemical separation have major disadvantages since they generate a concentrated stream that is more harmful than the original waste. In this study, our purpose is to reduce cutting oils by using biological treatment. Eighty one strains were isolated from cutting waste oil of industrial waste water sludge under aerobic conditions. Among these strains, KS47, which removed 90.4% cutting oil in 48 hr, was obtained by screening test under aerobic conditions(pH 7, $28^{\circ}C$). KS47 was identified as Pseudomonas aeruginosa according to morphological, physiological and biochemical properties, 16S rDNA sequence, and fatty acid analysis. P. aeruginosa KS47 could utilize cutting oil as carbon source. In batch test, we obtained optimal degradation conditions(1.5 g/L cell concentration, pH 7, and temperature $30^{\circ}C$). Under the optimal conditions, 1,060 mg/L cutting oil was removed 83.7% (74.1 mg/L/hr).

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.6
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• pp.1076-1081
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• 2001

Gamma-irradiated peanuts, Korean and Chinese origins, were investigated on detection properties by thermoluminescence (TL), electron spin resonance (ESR), and DNA comet assay (single cell gel electrophoresis). TL measurement showed that the non-irradiated sample revealed a glow curve with low intensity at about 25$0^{\circ}C$, while the irradiated samples showed higher intensity around at 18$0^{\circ}C$. TL ratio (TL$_1$/TL$_2$) of area for TL$_1$ glow curve to TL$_2$ was below 0.05 for the non-irradiated sample and 0.2 or more for the irradiated ones, thus identifying each other. ESR spectroscopy for the irradiated peanuts using outer skin showed negligible signals induced by irradiation, indicating ESR is little applicable to the detection of irradiated peanuts. In DNA comet assay, the non-sample had no or very short tails, whereas the irradiated samples revealed the cells with long tails. Significance in the increase of their lengths depending on irradiation dose (r=0.761/Korean, r=0.768/Chinese) was also found. There was no remarkable difference in detection properties by origins of samples in all determinations, It is concluded that TL analysis or DNA comet assay is suitable for detection of irradiated peanuts and a combined method is recommendable for enhancing the reliability of detection results.

• Molecules and Cells
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• v.40 no.12
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• pp.925-934
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• 2017

The Cdc6 protein is essential for the initiation of chromosomal replication and functions as a licensing factor to maintain chromosome integrity. During the S and G2 phases of the cell cycle, Cdc6 has been found to inhibit the recruitment of pericentriolar material (PCM) proteins to the centrosome and to suppress centrosome over-duplication. In this report, we analyzed the correlation between these two functions of Cdc6 at the centrosome. Cdc6 depletion increased the population of cells showing centrosome over-duplication and premature centrosome separation; Cdc6 expression reversed these changes. Deletion and fusion experiments revealed that the 18 amino acid residues (197-214) of Cdc6, which were fused to the Cdc6-centrosomal localization signal, suppressed centrosome over-duplication and premature centrosome separation. Cdc6 mutant proteins that showed defective ATP binding or hydrolysis did not exhibit a significant difference in suppressing centrosome over-duplication, compared to the wild type protein. In contrast to the Cdc6-mediated inhibition of PCM protein recruitment to the centrosome, the independence of Cdc6 on its ATPase activity for suppressing centrosome over-duplication, along with the difference between the Cdc6 protein regions participating in the two functions, suggested that Cdc6 controls centrosome duplication in a manner independent of its recruitment of PCM proteins to the centrosome.