• The Plant Pathology Journal
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• v.18 no.1
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• pp.1-5
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• 2002

The availability of nucleotide sequences of the coat protein gene of Potato leafroll virus (PLRV) permitted the construction of DNA primers that were utilized for cDNA synthesis. Polymerase chain reaction (PCR) products of a 487 bp. and approximately 500 bp DNA fragments were amplified from nucleic acid extracts of PLRV-infected tissue and strawberry mild yellow-edge (SMYE) diseased strawberry tissue, respectively. The amplified DNA fragments were further differentiated by hybridization analysis with a CDNA probe for the coat protein gene of PLRV and restriction fragment length polymorphism (RFLP) analysis. These results suggest that a luteovirus is associated with the SMYE disease.

• Animal cells and systems
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• v.3 no.1
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• pp.69-72
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• 1999

A tandemly repeated DNA sequence was identified and characterized y the combined RAPD and FISH data from a total genomic DNA of Welsh onion (Allium fistulosum). A clone containing this repeating sequence was selected and sequenced. This repeating unit of 314 bp inserted into pAf 072 contained 54.1% adenine and thymine residues, and showed the primer sequence used, 5'-GAAACGGGTG-3', in both terminals of the sequence. Fluorescence in situ hybridization using this tandemly repeated sequence as a probe indicated that the detected sites were coincident with the major C-banded constitutive heterochromatin in the terminal regions of both arms of all 6 chromosomes.

• Korean Journal of Microbiology
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• v.32 no.1
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• pp.47-52
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• 1994

The cells of Campylobacter jejuni heat-shocked at 48${\circ}C$ for 30 min synthesized the heat shock proteins of HSP90, HSP66 and HSP60. Those heat shock proteins were found to correspond to the heat shock proteins of HSP87, HSP66 (DnaK), and HSP58 (GroEL) of E. coli, respectively. By Southern blot analysis of the chromosomal DNAs of C. jejuni with groESL and dnaK genes of E. coli as DNA probes, the heat shock genes of C. jejuni which are homologous to the E. coli groESL and dnaK genes were found to exist in the chromosomal DNA. The genomic libraries of C. jejuni were constructed with the cosmid vector pWE15 and the groEL gene of C. jejuni were cloned in E. coli B178 groEL44 temperature senstive mutant. The hybrid plasmid (pLC1) was inserted with the DNA fragment (about 5.7kb in size) containing the groEL gene. E. coli groEL44 mutant cell transformed with the pLC1 could grow at 42${\circ}C$ by synthesizing the HSP60 of C. jejuni and regained the susceptibility to the ${\lambda}$ vir phage by expression of the groEL gene in the cloned cells. These indicated that the groEL products of C. jejuni had chaperon effects by synthesizing the heat shock proteins in the cloned cells of E. coli.

• Korean Journal of Veterinary Research
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• v.35 no.3
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• pp.531-536
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• 1995

Salmonella species are the most prevalent etiologic agents of food-borne acute gastroenteritis. Direct isolation of bacteria from the contaminated food, stool and animal tissues has been used for the diagnosis of salmonellosis routinely. However, isolation of bacteria is time consuming work and not so highly sensitive. In recent years, improved methods of polymerization chain reaction(PCR) and probe hybridization technique have led to the developement of diagnostic assays which employ to detect various human and animal pathogenic bacteria. In this study, we have performed the polymerization chain reaction to detect Salmonella pullorum from tissues and stool samples of chickens with two specific primers, ST5 and ST8C. The target DNA fragment of PhoE gene was successfully amplified from liver, spleen, pancreas, heart, lung, ovary, oviduct and feces samples. The amplified DNA fragments were hybridized with Salmonella typhymurium TA3000 PhoE probe by Southern hybridization. The PCR to amplify the PhoE gene was highly rapid and sensitive method to detect Salmonella pullorum from tissues and stool samples.

• Korean Journal of Ecology and Environment
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• v.54 no.3
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• pp.209-220
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• 2021

Pacific herring, Clupea pallasii, a keystone species with significant ecological and commercial importance, is declining globally throughout much of its range. While traditional fishing equipment methods remain limited, new sensitive and rapid detection methods should be developed to monitor fisheries resources. To monitor the presence and quantity of C. pallasii from environmental DNA (eDNA) extracted from seawater samples, a pair of primers and a TaqMan^® probe specific to this fish based on mitochondrial cytochrome b (COB) sequences were designed for the real-time PCR (qPCR) assay. The combination of our molecular markers showed high specificity in the qPCR assay, which affirmed the success of presenting a positive signal only in the C. pallasii specimens. The markers also showed a high sensitivity for detecting C. pallasii genomic DNA in the range of 1 pg~100 ng rxn^-1 and its DNA plasmid containing COB amplicon in the range of 1~100,000copies rxn^-1, which produced linear standard calibration curves (r²=0.99). We performed a qPCR assay for environmental water samples obtained from 29 sampling stations in the southeastern coastal regions of South Korea using molecular markers. The assay successfully detected the C. pallasii eDNA from 14 stations (48.2%), with the highest mean concentration in Jinhae Bay with a value of 76.09±18.39 pg L^-1 (246.20±58.58 copies L^-1). Our preliminary application of molecular monitoring of C. pallasii will provide essential information for efficient ecological control and management of this valuable fisheries resource.

• Proceedings of the Korean Biophysical Society Conference
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• 2002.06b
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• pp.57-57
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• 2002

Junctate is a newly identified integral endo(sarco)plasmic reticulum membrane calcium binding protein, which is an alternative splicing form of the same gene generating aspartyl-hydroxylase and junctin. Screening a mouse heart cDNA library using canine junctin cDNA as a probe yielded 3 complete mouse heart cDNAs.(omitted)

• Korean Journal of Veterinary Research
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• v.33 no.1
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• pp.43-51
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• 1993

Mitochondrial DNA(mt DNA) of Mammalian is the circular one which the 16.5K base pairs and show the maternal inheritance. Evolutional speed of nucleotide sequence is very fast. So that polymorphic analysis of mt DNA provide the useful informations to investigate the genetic relations of interspecies. Authors trials were focussed to compare with the polymorphic differences of mitochondrial DNA between Jindo and Japanese mongrel dogs. DNA was extracted from bloods of 21 head of Jindo dogs and 20 head of Japanese dogs and isolated using 10 kinds of restriction endonucleases(Apa I, BamH I, Bgl II, EcoR I, EcoR V, Hinc II. Hind III, Pst I, Sty I, Xba I) and then separated by the agarose gel electrophoresis. After sourthern blotting hybridization was completed using the mtDNA of Japanese mongrel dogs as a probe. Autoradiography was used to compare the polymorphism of mtDNA both dogs. The results obtained were as follows; 1. mt DNA of Jindo dog showed polymorphism resulting cleavage with four kinds of restriction endonuclease, Apa I, EcoR V, Hinc II, Sty I. While in the Japanese mongrel dogs observed the polymorphism in the five kinds of restriction endonuclease supplemented with EcoR I. 2. Compared with both dogs the frequency differences of DNA polymorphism were recognized in the specific restriction endonuclease Apa I. Consequently in the restriction endonuclease Apa I both dogs classified with three types as A, B, C however in the Jindo dogs frequency of C type was 71.5 percent but in Japanese mongrel dogs observed 45 percent in the A type. 3. DNA polymorphism obtained from the use of five kinds of restriction endonuclease were classified with seven types. In Jindo dogs frequency was highest in the type 6 as 71.4 percent but in the Japanese mongrel dogs showed 35 percent in the type 5. 4. Genetic distances calculated by NEI method showed 0.0089 in Jindo dogs and was 0.0094 in the Japanese mongrel dogs.

• BMB Reports
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• v.35 no.3
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• pp.320-329
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• 2002

A gene, coined tay, for a thermostable DNA polymerase from the novel, extremely thermophilic bacterium Thermoanaerobacter yonseiensis was cloned and expressed in E. coli. Using a DNA polymerase homologous PCR product as a hybridization probe, tay was isolated and sequenced to consist of 2621 nucleotides that encode 872 amino acids. A database analysis showed that DNA polymerase, coined Tay, from T. yonseiensis shared a 39% to 47% identity in the amino acid sequence with those from other DNA polymerases. Tay was overexpressed in E. coli as a fusion protein with a poly-histidine tag at the C-terminus. It was purified by heat treatment, followed by a $Ni^{2+}$-chelate column. The molecular weight of purified Tay was approximately 97 kDa, as shown by SDS PAGE, and it showed high DNA polymerase activity and thermostability. However, it had no 3'$\rightarrow$5' exonuclease activity.

• Journal of Life Science
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• v.6 no.4
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• pp.286-292
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• 1996

Angiogenesis is a fundamental process by which new blood vessels are formed. It is essential in embryo development, and wound healing. Furthermore, malignant tumor growth and metastasis are also angiogenesis-dependent. In the catilage tissue, normal angiogenesis process is suppressed. In fact, it was reported that angiogenesis-inhibitory substances were isolated from the extracts of cow and shark catilage tissue. In order to isolate genes involved in the regulation of angiogenesis from a catilage fish, we constructed a shark cDNA library from Scylliohinus torazame. We then screened the library using hyman tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) gene as a probe. Among the 4 X 10$^{4}$ plaques screened, we isolated 2 positive clones (T-1, T-2). Restriction enzyme analysis revealed that the T-1 clone contains 0.8 kb cDNA insert, and the T-2 clone contains 1.2 kb and 2.2 kb inserts, respectively. Further DNA sequence analysis shows that the DNA sequence of the T-1 clone is 53% homologous to that of the human TIMP-1 gene.

• The Korean Journal of Nuclear Medicine
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• v.37 no.1
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• pp.43-52
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• 2003

Microarray technology allows the simultaneous analysis of gene expression patterns of thousands of genes, in a systematic fashion, under a similar set of experimental conditions, thus making the data highly comparable. In some cases arrays are used simply as a primary screen loading to downstream molecular characterization of individual gene candidates. In other cases, the goal of expression profiling is to begin to identify complex regulatory networks underlying developmental processes and disease states. Microarrays were originally used with ceil lines or other simple model systems. More recently, microarrays have been used in the analysis of more complex biological tissues including neural systems and the brain. The application of cDNA arrays in neuropsychiatry has lagged behind other fields for a number of reasons. These include a requirement for a large amount of input probe RNA In fluorescent-glass based array systems and the cellular complexity introduced by multicellular brain and neural tissues. An additional factor that impacts the general use of microarrays in neuropsychiatry is the lack of availability of sequenced clone sets from model systems. While human cDNA clones have been widely available, high qualify rat, mouse, and drosophilae, among others are just becoming widely available. A final factor in the application of cDNA microarrays in neuropsychiatry is cost of commercial arrays. As academic microarray facilitates become more commonplace custom made arrays will become more widely available at a lower cost allowing more widespread applications. in summary, microarray technology is rapidly having an impact on many areas of biomedical research. Radioisotope-nylon based microarrays offer alternatives that may in some cases be more sensitive, flexible, inexpensive, and universal as compared to other array formats, such as fluorescent-glass arrays. In some situations of limited RNA or exotic species, radioactive membrane microarrays may be the most practical experimental approach in studying psychiatric and neurodegenerative disorders, and other complex questions in the brain.