• Journal of Animal Science and Technology
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• v.44 no.1
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• pp.13-22
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• 2002

To identify the differentially expressed genes at growth stage of Hanwoo, we constructed the subtractive cDNA library from loin mRNA of 12- and 24-month old Hanwoo by PCR-based subtraction. The fourteen genes were confirmed by sequencing and reverse northern blot analysis, and they were selected as candidate of putative genes differentially expressed at the growth stage of Hanwoo. Three subtracted cDNA fragments that expressed specific signal to cDNA probe for 6-month-old loin of Hanwoo were highly homologous to those of the genes encoding EPV 20, Ca2＋ATPase, and TCTP, respectively. The nine cDNA clones showed intense signal to cDNA probe from 12-month-old loin of Hanwoo, and highly homologus to those of genes encoding VCP, HSP 70, aldolase A, MSSK1, GM-2 activator protein, ryanodine receptor, acidic ribosomal phosphoprotein p1, ADP/ATP translocase, and UCP 2, respectively. Two subtracted cDNA clones that expressed specific signal to cDNA probes for 12- and 24-month-old loin of Hanwoo were detected. One of them was highly homologus to the gene encoding ferrochelatase and the other was highly homologus to the gene encoding ADRP.

• KIEE International Transactions on Electrophysics and Applications
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• v.4C no.4
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• pp.133-136
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• 2004

This research aims to develop a DNA chip array without an indicator. We fabricated a microelectrode array through photolithography technology. Several DNA probes were immobilized on an electrode. Then, target DNA was hybridized and measured electrochemically. Cyclic-voltammograms (CVs) showed a difference between the DNA probe and mismatched DNA in an anodic peak. This indicator-free DNA chip resulted in a sequence-specific detection of the target DNA.

• Korean Journal of Veterinary Research
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• v.34 no.2
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• pp.387-394
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• 1994

To make the genomic DNA probe of Theileria sergenti, the merozoites were purified from bovine erythrocytes. The infected erythrocytes were lysed by Aeromonas hydrophila(Ah-1) hemolysin, and the parasites were isolated by ultracentrifugation on a Percoll discontinuous density gradient. For construction of a T sergenti genomic DNA library, T sergenti DNA was digested with Pstl and the fragments were ligated into the PstI site of pUC19 before transformation of Escherichia coli JM83. Out of thousands of transformants obtained by transformation of E coli JM83 with the genomic library, three plasmids were chosen. The sizes of the inserted DNAs were 2.9kb(2.4kb and 0.5kb) in pKTS1, 4.3kb in pKTS2 and 1.5kb in pKTS3, respectively. The DNA fragments used as probe KTS1(2.4kb), KTS2(4.3kb) and KTS3(1.5kb) were labeled digoxigenin-11-dUTP for the Southern hybridization. In Southern hybridization, all of the probes(KTS1, KTS2 and KTS3) reacted specifically to T sergenti DNA, but not to bovine leucocyte DNA. In order to find out the sensitivities of the digoxigenin-11-dUTP-labeled KTS1 and KTS3 as the probes, purified merozoite DNA and bovine DNA (control) were checked by dot blot hybridization with the probes. Both of the probes, KTS1 and KTS3, detected as minimum amount of 975pg of the T sergenti DNA, but not bovine DNA even to 500ng.

• The Plant Pathology Journal
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• v.37 no.3
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• pp.291-298
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• 2021

False smut caused by Ustilaginoidea virens is an important rice fungal disease that significantly decreases its production. In the recent past, conventional methods have been developed for its detection that is time-consuming and need high-cost equipments. The research and development in nanotechnology have made it possible to assemble efficient recognition interfaces in biosensors. In this study, we present a simple, sensitive, and selective oxidized graphene-based geno-biosensor for the detection of rice false smut. The biosensor has been developed using a probe DNA as a biological recognition element on paper electrodes, and oxidized graphene to enhance the limit of detection and sensitivity of the sensor. Probe single-stranded DNA (ssDNA) and target ssDNA hybridization on the interface surface has been quantitatively measured with the electrochemical analysis tools namely, cyclic voltammetry, and linear sweep voltammetry. To confirm the selectivity of the device, probe hybridization with non-complementary ssDNA target has been studied. In our study, the developed sensor was able to detect up to 10 fM of target ssDNA. The paper electrodes were employed to produce an effective and cost-effective platform for the immobilization of the DNA and can be extended to design low-cost biosensors for the detection of the other plant pathogens.

• Microbiology and Biotechnology Letters
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• v.15 no.2
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• pp.116-121
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• 1987

In order to cloning of the nif genes of Enterobacter agglomerans NFB-264, the digested total DNA of the strain was ligated to pBR 322 and transformed into E. coli. Through the negative selection and colony hybridization, the transformants were obtained. The recombinant plasmids, pNEL 10 and pNES 20 were extracted from these transformants. It was known from Southern hybridization that pNEL 10 contained the 12 Mdal foreign DNA fragment hybridized with nif Q-X probe and pNES 20 included the 5 Mdal foreign DNA fragment hybridized with nif NE and nif YK probe.

• Proceedings of the Korean Information Science Society Conference
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• 2006.10a
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• pp.516-521
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• 2006

DNA칩의 성능은 칩을 구성하는 probe에 의해 결정된다. 좋은 probe는 homogeneity, sensitivity, specificity와 같은 속성을 갖추어야 한다. 이중 specificity는 probe의 특정 유전자에 대한 선별적 결합 능력을 나타내는 것으로 이를 계산하는데 가장 많은 시간이 요구된다. 본 논문은 유전자의 개별 후보 probe들에 대한 선정 작업을 실행하기 전에 q-gram을 이용하여 비교가 필요한 유전자들만을 전체 유전자의 길이가 n인 경우 O(1/$4^an^2$)의 시간에 선별하는 전처리 알고리즘을 제안한다. 그리고 제안한 알고리즘을 사용함으로써 기존 방법들보다 빠른 probe 선정이 가능함을 실제 데이터를 사용하여 보인다.

• 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.

• Korean Journal of Veterinary Research
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• v.33 no.3
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• pp.479-486
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• 1993

This study was attempted co develope a method for detection of Theileria sergenti infection on the basis of hybridization of parasite DNA with a probe. For construction of a T sergenti genomic library, T sergenti DNA was digested completely with Bam-HI and the fragments were ligated into the Bam-HI site of pUC-19 before transformation of Escherichia colistrain JM83. To detect clones containing the parasite's DNA sequences, a genomic DNA library of T sergenti constructed in pUC-19 was screened by cracking and Southern hybridization. Seven colonies were chosen from 29 colonies which were screened by transformation of Escherichia coli strain JM83. Seven transformants were comfirmed from seven colonies by cracking. The sizes of transformants were about 5Kb, 5.7Kb, 4.3Kb, 7.75Kb, 7.85Kb, 5.8Kb, 3.8Kb, respectively. DNA inserts, T sergenti DNA, and bovine DNA were hybridized with radio-labelled T sergenti DNA. Two($pT_1$, $pT_1$) of the seven inserts and T sergenti DNA reacted strongly but another 5 inserts and bovine DNA showed weak reation. All of the DNA inserts were not reaction, but T sergenti DNA were very weakly and bovine DNA were strongly reacted to hybridization with radio-labelled bovine DNA. Therefore, we obtained total 7 T sergenti DNA fragments in this study.

• Journal of Life Science
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• v.13 no.2
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• pp.163-167
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• 2003

Identification of species within the toxin-producing genus Alexandrium is vital for biotoxin monitoring and mitigation decisions regarding shellfish industry. In particular, the discrimination of resting cysts of only A. tamarense from that of Alexandrium spp. is considerable important to fundamentally monitor and predict this species before vegetative cells occur in the nature. Fluorescent cTAM-F1 DNA probe was responsible to not only binding the activity of the vegetative cells in A. tamarense, but also to the resting cysts, which was treated with methanol after fixation and stained by primuline on the surface The location of fluorescence in cultured vegetative cells and resting cysts was almost at tile bottom of the nucleus. The optimal incubation temperature and time using in situ hybridization were 50-$54^{\circ}C$ and 40-60 min, respectively, to penetrate the DNA probe into cell.

• JSTS:Journal of Semiconductor Technology and Science
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• v.1 no.4
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• pp.232-238
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• 2001

A new DNA purification chip is proposed and fabricated for the sample preparation of Nucleic Acid (NA) probe assay. The proposed DNA purification chip is fabricated using photosensitive glass substrate and polydimethylsiloxane (PDMS) cover fixture. We have successfully captured and eluted the DNA using the fabricated photosensitive glass chip. The fabricated DNA purification chip showed a binding capacity of $15ng/\textrm{cm}^2$and a minimum extractable input concentration of $100copies/200\muL$. The proposed DNA purification chip can be applied for low-cost, disposable sample preparation of NA probe assays.