• Journal of Korean society of Dental Hygiene
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• v.6 no.4
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• pp.311-324
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• 2006

The purpose of this investigation was to evaluate of the specificity of Fusobacterium nucleatum subspecies-specific DNA probes using dot blot hybridization. To confirm whether the clinical isolates were F. nucleatum or not, 16S rDNA of them were cloned and sequenced. The sequencing data were used in homology search with database of GenBank. When the homology was above 98% compared with the nucleotide sequence of a certain bacteria, it was judged as the same species with the bacteria. 23 strains of F. nucleatum were isolates from subgingival plaque of periodontitis patient. The clinical isolates of F. nucleatum were classified into 10 groups using phylogenetic analysis of 16S rDNA sequence. F. nucleatum subspecies nucleatum-specific DNA probe Fu4(1.3 kb) reacted with genomic DNAs from 8 type strains of F. nucleatum and it reacted strongly with those from 8 clinical isolates. The Fp4(0.8 kb) reacted with F. nucleatum subsp. polymorphum ATCC 10953 and one clinical isolates. Fv35(1.9 kb) and Fs17(8.2 kb) probes reacted with genomic DNAs from F. nucleatum subsp. vincentii ATCC 49256 and F. nucleatum subsp. fusiform ATCC 51190, respectively. Our results showed that it is not enough to evaluate the specificity of F. nucleatum subspecies-specific DNA probes with only dot blot hybridization. Therefore, Southern blot analysis will be necessary to confirm the specificity of F. nucleatum subspecies-specific DNA probes.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.859-871
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• 1996

This study was conducted to elucidate the distribution of Aujeszky's disease viral nucleic acids and antigens in the central nervous system (CNS) of piglets. The first Korean isolate of Aujeszky's disease virus(ADV) that isolated from naturally infected piglets in Yang San, was inoculated into 32 day old piglets with $10^{5.9}TCID_{50}/ml$ through intranasal or intramuscular route. These piglets were sacrificed at every 24hrs for 8 days. The immunohistochemistry (IHC) was conducted to detect the viral antigens in paraffin-embedded tissue sections using avidin-biotin-peroxidase complex (ABC) method. The viral nucleic acids were detected by in situ hybridization (ISH) using ADV specific DNA probe labeled with digoxigenin. The ADV antigens were detected in reticuloendothelial cells of spleen, lymph nodes and tonsil, alveolar walls, leptomeningeal vascular walls, inflammatory foci of each organ, and nerve cells. The viral nucleic acids were detected in the spinal trigeminal nucleus and its tracts of the pons and medulla oblongata by the ISH technique. The pathways of AD viruses in CNS were determined by IHC and ISH. In the intranasally inoculated group, the viruses in nasal mucosa moved to medulla oblongata and pons through the trigeminal nerve. In case of intramuscullarly inoculated group, viruses moved to brain via lymphoid organs or spinal nerves from sciatic nerves.

• The Korean Journal of Malacology
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• v.29 no.3
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• pp.181-187
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• 2013

In order to investigate environmental stress inducible genes in abalone, we analyzed differentially expressed transcripts from a Pacific abalone, Haliotis discus hannai, after exposure to heat-, cold- or hyposalinity-shock by suppression subtractive hybridization (SSH) method. 1,074 unique sequences from SSH libraries were composed to 115 clusters and 986 singletons, the overall redundancy of the library was 16.3%. From the BLAST search, of the 1,316 ESTs, 998 ESTs (75.8%) were identified as known genes, but 318 clones (24.2%) did not match to any previously described genes. From the comparison results of ESTs pattern of three SSH cDNA libraries, the most abundant EST was different in each SSH library: small heat shock protein p26 (sHSP26) in heat-shock, trypsinogen 2 in cold-shock, and actin in hyposalinity SSH cDNA library. Based on sequence similarities, several response-to-stress genes such as heat shock proteins (HSPs) were identified commonly from the abalone SSH libraries. HSP70 gene was induced by environmental stress regardless of temperature-shock or salinity-stress, while the increase of sHSP26 mRNA expression was not detected in cold-shock but in heat-shock condition. These results suggest that the suppression subtractive hybridization method is an efficient way to isolate differentially expressed gene from the invertebrate environmental stress-response transcriptome.

• Journal of Sericultural and Entomological Science
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• v.50 no.2
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• pp.122-125
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• 2012

This study was aimed for a development of a useful gene promoter which has a transcript expressional specificity in the early embryonic period of the silkworm, Bombyx mori. To select a useful gene expressed in the early embryonic stage, we constructed and analyzed a PCR-base subtraction cDNA library. In subtractive hybridization analysis, we confirmed four clones as differently expressed genes(BmNanos-like, BmNanos-P, BmNanos-O, BmVasa mRNAs). Northern hybridization and real time PCR results reveled that the BmNanos-like gene promoter is suitable for the silkworm transgenic vector system. Further defined studies on molecular functions and biological roles of their promoters will give us well-fined information and its application.

• The Journal of the Acoustical Society of Korea
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• v.26 no.1
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• pp.42-47
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• 2007

In this paper. we have studied improvement in sensitivity by increasing the frequency of SAW sensors for detecting the immobilization and hybridization of DNA. The sensor consists of twin SAW delay lines operating at 200MHz, a sensing channel and a reference channel. fabricated on $36^{\circ}$ rotated Y-cut X-propagation $LiTaO_3$ crystals. The optimum concentration of probe and target DNA was decided for the improvement of detection mechanism. and digital syringe pump system was used to reduce the human errors. The hybridization between immobilized probe DNA and target DNA on the gold-coated delay line results in mass loading on the delay line of the sensing channel. Thus, the relative frequency change was monitored in relation to the mass loading. The measurement results showed a good response of the sensor to the DNA hybridization with a maximum sensitivity level up to 0.066ng/m1/Hz.

• Journal of the Korean Society for Precision Engineering
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• v.29 no.12
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• pp.1304-1312
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• 2012

According to the successful development of hybrid vehicle, hybridization of construction equipments like excavator, wheel loader, and backhoe etc., is gaining increasing attention. However, hybridization of excavator and commercial vehicle is very different. Therefore a specialized energy management control algorithm for excavator should be developed. In this paper, hybridization of excavators is investigated and a new energy management control algorithm is proposed. Four control parameters, i.e., lower baseline, upper baseline, idling generation speed, and idling generation torque, are newly introduced and a new operating principle using those four control parameters is proposed. The use of Genetic Algorithm for the optimization of the four control parameters from the view point of minimization of fuel consumption for standard excavating operation is suggested. In order to verify the proposed algorithm, dedicated simulation program of hybrid excavator was developed. The proposed algorithm is applied to a specific hydraulic excavator and 20.7% improvement of fuel consumption is achieved.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.5
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• pp.603-609
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• 2011

Telomeres are special structures at the ends of eukaryotic chromosomes. Vertebrate telomeres consist of tandem repeats of conserved TTAGGG sequence and associated proteins. Birds are interesting models for molecular studies on aging and cellular senescence because of their slow aging rates and longer life spans for their body size. In this longitudinal study, we explored the possibility of using telomeres as an age-marker to predict age in Single Comb White Leghorn layer chickens. We quantified the relative amount of telomeric DNA in isolated peripheral blood lymphocytes by the Quantitative Fluorescence in situ Hybridization technique on interphase nuclei (IQ FISH) using telomere-specific DNA probes. We found that the amount of telomeric DNA (ATD) reduced significantly with an increase in chronological age of the chicken. Especially, the telomere shortening rates are greatly increased in growing individuals compared to laying and old-aged individuals. Therefore, using the ATD values obtained by IQ FISH we established the possibility of age prediction in chickens based on the telomere theory of aging. By regression analysis of the ATD values at each age interval, we formulated an equation to predict the age of chickens. In conclusion, the telomeric DNA values by IQ FISH analyses can be used as an effective age-marker in predicting the chronological age of chickens. The study has implications in the breeding and population genetics of poultry, especially the reproductive potential.

• The Plant Pathology Journal
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• v.33 no.1
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• pp.80-86
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• 2017

One of the major problems in strawberry production is difficulty in diagnosis of anthracnose caused by Colletotrichum acutatum or Glomerella cingulata in latent infection stage. We here developed a diagnostic tool for the latent infection consisting of initial culturing of fungi, DNA extraction, synthesis of PCR-amplified probes and microtube hybridization (MTH) using a macroarray. The initial culturing step is convenient to lure the fungi out of the plant tissues, and to extract PCR-inhibitor-free DNA directly from fungal hyphae. For specific detection of the fungi, PCR primers were designed to amplify the fungal MAT1-2 gene. The subsequent MTH step using the PCR products as probes can replace the laborious electrophoresis step providing us sequence information and high-throughput screening. Using this method, we have conducted a survey for a few thousands nursery plants every year for three consecutive years, and finally succeeded in eliminating latent infection in the third year of challenge.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.282-285
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• 2001

The microbial community structure and in situ spatial distribution of ammonia oxidizing and nitrite oxidizing bacteria in nitrifying biofilm of an upflow biological aerated filter system were investigated. The reactor had been continuously operated under high free ammonia concentration and low DO concentration for nitrite accumulation more than 2 years before the experiment. Fluorescence in situ hybridization

• Molecules and Cells
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• v.23 no.1
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• pp.100-107
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• 2007

The moss Physcomitrella patens has two life cycles, filamentous protonema and leafy gametophore. A modified from of suppression subtractive hybridization (SSH), mirror orientation selection (MOS), was applied to screen genes differentially expressed in the P. patens protonema. Using reverse Northern blot analysis, differentially expressed clones were identified. The identified genes were involved mainly in metal binding and detoxification. One of these genes was an AP2 (APETALA2) domain-containing protein (PpACP1), which was highly up-regulated in the protonema. Alignment with other AP2/EREBPs (Ethylene Responsive Element Binding Proteins) revealed significant sequence homology of the deduced amino acid sequence in the AP2/EREBP DNA binding domain. Northern analysis under various stress conditions showed that PpACP1 was induced by ethephon, cadmium, copper, ABA, IAA, and cold. In addition, it was highly expressed in suspension-cultured protonema. We suggest that PpACP1 is involved in responses to metals, and that suspension culture enhance the expression of genes responding to metals.