• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1976.10a
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• pp.187.4-188
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• 1976

서로 다른 11주의 Bacillus coagulans와 13종의 Bacillus 속 14주를 deoxyribonucleic acid (DNA) -DNA hybridization method에 의해서 분류학적인 연구를 하였다. 사용한 B. coagulans 11주종 6주는 흙에서(일본 오사까교회) 분리했고 나머지 5주는 ATCC, IFO에서 authentic strains을 얻어서 사용했다. 사용된 B. coagulans는 Bergey's Manual(8 thed)에 의거 Grodon씨들의 방법으로 동정한 결과 B. coagulans로서 확인되었다.(중략)

• Proceedings of the KIEE Conference
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• 2004.07c
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• pp.2128-2130
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• 2004

High throughput analysis using a DNA chip microarray is powerful tool in the post genome era. Less labor-intensive and lower cost-performance is required. Thus, this paper aims to develop the multi-channel type label-free DNA chip and detect SNP (Single nucleotide polymorphisms). At first, we fabricated a high integrated type DNA chip array by lithography technology. Various probe DNAs were immobilized on the microelectrode array. We succeeded to discriminate of DNA hybridization between target DNA and mismatched DNA on microarray after immobilization of a various probe DNA and hybridization of label-free target DNA on the electrodes simultaneously. This method is based on redox of an electrochemical ligand.

• Proceedings of the KIEE Conference
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• 2007.07a
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• pp.1375-1376
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• 2007

A synthesized 21-mer single-stranded DNA(ss DNA) was covalently immobilized onto a self-assembled aminoethanethiol monolayer modified gold electrode onto QCM. The covalently immobilized ssDNA was hybridized with complementary ssDNA. The interaction between surface immobilized ssDNA and complementary 21-mer DNA in solution was also examined. Each step was followed by monitoring changes in the QCM frequency with time. Also, PBS with pH 7.0 was selected as a supporting electrolyte in order to get maximum sensitivity and good bioactivity.

• 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.

• 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.

• Proceedings of the KIEE Conference
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• 2002.11a
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• pp.260-262
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• 2002

The determination of DNA hybridization can apply the molecular biology research. clinic diagnostics. bioengineering, environment monitoring, food science and other application area. So, The determination of hybridization is very important for the improvement of DNA detection system. In this study, we report the characterization of the DNA hybridization by the electricalchemical methods. The probe oligonucleotide was used to determine the amount of target oligonucleotide in solution using Methylen Blue(MB) as the electrochemical indicators. The cathodic peak currents($I_{peak}$) of MB were linearly related to the concentration of the target oligonucleotide sequence in the range $1[{\mu}M]{\sim}0.1[{\mu}M]$. The detection limit of this approach was 0.01[nM]. As a result, the match oligonucleotide(CR-1) was most stable state and the peak of redox current measured by DNA hybridization detection sensors by using electrochemical method seem to be similar to 1-mer terminal mismatch oligonucleotide(MR-3). The MR-2, MR-3, MR-22 and MR-33 have each mismatching sequence of central and terminal. With this set the role of point mutations was to be investigated. Terminal mismatch oligonucleotide (MR-3, 33) is shown more stable state than central mismatch oligonucleotide(MR-2, 22). And 1-mer mismatch oligonucleotide(MR-2 or 3) is shown more stable state than 2-mer mismatch oligonucleotide(MR-22 or 33).

• Microbiology and Biotechnology Letters
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• v.20 no.3
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• pp.280-288
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• 1992

Molecular cloning of metalloprotease gene from Serratia marcescens ATCC 21074 into Escherichia coli JM109 was carried out. Chromosomal DNA of S. marcescens was completely digested with Hind111 and southern hybridization with a synthetic oligonucleotide probe revealed that a 50 KD metalloprotease gene was contained in 4.0 Kb chromosomal DNA fragment, 4.0 Kb chromosomal DNA fragments eluted from agarose gel were ligated with pUC19 and transformed into E. coli JM109. Nine positive clones were obtained from about $1\times 10^3$ transformants by colony hybridization. Their recombinant plasmids, pSPl and pSP2 have same chromosomal DNA fragments in pUC19 in opposite-orientations. When cloned metalloprotease gene was expressed in E. coli, about 52 KD precursor protein of metalloprotease was detected by western blot analysis from E. coli harboring a recombinant plasmid pSP2. Plasmid pSP2 showed no protease activities in E. coli but overproduced the active metalloprotease in S. rnarcescens ATCC 27117.

• The Korean Journal of Mycology
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• v.26 no.2 s.85
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• pp.135-143
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• 1998

Contour-clamped homogeneous electric field gel electrophoresis was used to establish electrophoretic karyotype for 10 species of Fusarium sections Sporotrichiella, Liseola, Gibbosum, Discolor and Martiella. Intact chromosomal DNA was isolated from fungal protoplast and separated under various conditions according to their size in order to improve DNA separation. The numbers of chromosome-sized DNA molecules for individual species ranged from 5-13, with individual chromosomes ranging from 0.78 Mb to 7.20 Mb in size. The total genome DNA size of each species was estimated at about 18.32 Mb to 48.20 Mb. Comparison of karyotype profiles following Southern hybridization analysis with a randomly selected genomic probe of F. oxysporum formae speciales litii was carried out.

• Microbiology and Biotechnology Letters
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• v.20 no.4
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• pp.483-490
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• 1992

In order to understand the transfer behavior of a particular gene in water environments, kanamycin resistance ($Km^r$) gene was tracked by Southern hybridization with DNA probe in its conjugal transfer. A $Km^r$ natural bacterial isolate and genetically modified microorganisms (GMMs) constructed from the isolate were used as donor for conjugal transfer of the $Km^r$ gene. The transfer frequencies of the $Km^r$ gene from GMM strains were generally 10 to 100 times higher than those from the natural isolate. The conjugants obtained from GMM strains in LB broth had more plasmids newly appeared, and particularly the conjugants in A Wand FW waters revealed more rearrangement in their plasmids as a function of conjugation time. When plasmids of the conjugants obtained in LB broth were Southern hybridized with DNA probe of the $Km^r$ gene, the $Km^r$ plasmids in the conjugants were detected at the same position of the plasmids in donor cells, in spite of the fact that the plasmids were highly rearranged in conjugant cells. But the $Km^r$ plasmids in the donor of DKI and DKC601, and DKC600 were not identified in the conjugants obtained after 50 h conjugation in AW and after 30 h in AW, respectively. The size of the $Km^r$ plasmids showing hybridization signal were a little changed in the conjugants obtained in A Wand FW waters. Therefore, the method of Southern hybridization with DNA probe was proved to be very specific and useful for tracking of particular genes in water environments.

• YAKHAK HOEJI
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• v.33 no.5
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• pp.300-307
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• 1989

Expression of transferrin gene in various organs of rat was studied using rat transferrin cDNA. The hybridization method of $[^{35}S]-labeled$ transferrin cDNA with transferrin mRNA in cytoplasmic preparations was used to measure the level of transferrin mRNA. The rat from 15-day old fetus to 21-day old postnatal were employed as an animal model. In the liver, the level of transferrin mRNA increased with increasing age. However, the level of transferrin mRNA in brain was significantly lower than that in liver and the level did not increase with age.