• Parasites, Hosts and Diseases
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• v.34 no.3
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• pp.191-196
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• 1996

Polymerase chain reaction (PCR) is a powerful technique to detect scanty amount of DNA from living organisms. The present study intended to develope specific primers for PCR diagnosis of pneumocystosis and to evaluate diagnostic efficacy by preparation of template DNAs from invasive BAk fluid and also to screen serum or blood as a non-invasive specimen. Albino rats of Wistar or Fischer strains were experimentally infected by Pneumocwstis ccrinii. Extracted DNAs or cell Iysates of their blood, bronchoalveolar lavage fluid, and lung homogenate were used as the tenlplate DNA. Primers were synthetic oligonucleotides among 16s rDNA sequences. All of the primer combinations gave PCR products, but the primer pair of #24 and #27 gave best quality product of 666 bp. The sensitivity of PCR with Iysates of BAk fluid was 57.7% but it increased to 84.6% with extracted DNAs. None of BAL Iysate or DNA was positive among 13 microscopically negatives. The serum DNAs were positive only in 2 cases out of 20 morphologically positive rats. DNAs of human, rat, other parasites, yeast, and microorganisms were negative. The findings suggest that the present primers are specific but simple Iysate of BAL fluid is not sensitive. PCR may be used as a routine diagnostic method of pneumocystosis if simple and rapid preparation of non-invasive clinical specimens are available.

• Journal of Korean Society of Forest Science
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• v.83 no.1
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• pp.20-24
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• 1994

In woody species with a long life span, the studies on inheritance of any trait may be very time consuming and laborious. Chloroplast DNA(cpDNA) has been a valuable tool in such studies since it has several unique features such as limited genome size and cytoplasmic inheritance. In the present study, cpDNAs from five different species of Populus(P. alba, P. glandulosa, P. alba${\times}$P. glandulosa, P. davidiana, and P. nigra), and Nicotiana tabacum were compared with regard to restriction fragment length polymophism. The results showed that cpDNAs among the species were very conserved, although some polymorphisms were observed when the DNAs were digested with restriction enzyme EcoRI or KphI. The other enzymes (Bgl II, and PstI) tested produced identical restriction fragmentation pattern among the species. However, cpDNAs from all the five Populus species showed different restriction fragmentation pattern from that of tobacco with the four restriction enzymes tested. Southern hybridization with tobacco rbcL gene fragment as a probe also produced identical pattern among Populus species. The results indicate that cpDNAs in the genus are very well conserved during evolution.

• Korean Journal of Veterinary Research
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• v.42 no.3
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• pp.363-370
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• 2002

For the development of diagnostic polymerase chain reaction (PCR) to fungal infection by dermatophytes Trichophyton and Microsporum, detection of the fungal DNA by PCR and analysis of the DNA pattern were undertaken in the present study. A total of 15 strains were tested and those consisted of 3 reference strains and 12 isolates such as: reference strains of T mentagrophytes (downy type, ATCC 9533), T rubrum (IFO 6204) and M gypseum (ATCC 9083), and each isolate of T mentogrophytes (powdery type), T mentagrophytes (granular type), T mentogrophytes (purple-red type), T rubrum, T raubitschekii, T tonsurans, T equinum, T ajelloi, T verrucosum, M cookei, M nanum and M gypseum. The DNA were purely isolated from all strains of Trichophyton spp. and Microsporum spp. by a simple method partly consisted of disruption of fungal cells by lyophilization and grinding and extraction of fungal DNA without phenol treatment which is a routine procedure in DNA isolation. For the detection of fungal DNAs, optimal condition of PCR was determined as preheating once at $94^{\circ}C$ for 5 min, 35 cycles of denaturation at $94^{\circ}C$ for 1 min, annealing at $38^{\circ}C$ for 1 min and polymerization at $72^{\circ}C$ for 2 min, and 1 cycle of final extension at $72^{\circ}C$ for 5 min. In PCR using arbitrary primers AP-1 (5' ACCCGACCTG3') and AP-2 (5' ACGGGCCAGT3'), DNAs in various numbers and sizes were detected from different species of Trichophyton and Microsporum, while DNAs in similar size were also detected in all strains of Trichophyton spp. and Microsporum spp. There were unique DNAs observed from certain dermatophytes by AP-1 such as 1,900 bases in T rubrum, 950 and 1,100 bases in T raubitscheldi, 2,100 bases in T equinum, 400 bases in T verrucosum and 1,150 bases in M gypseum. The unique DNAs were also observed by AP-2 such as 1,200 bases in T ajelloi, 250 bases in T verrucosum, 1,150 bases in M cookei and 2,000 bases in M nanum. The results indicated that PCR can detect a specific DNA from certain Trychophyton and Microsporum spp, which can be the information for further development of diagoomc PCR to dennatophytes.

• Journal of Plant Biotechnology
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• v.37 no.2
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• pp.174-185
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• 2010

Full-length cDNAs are essential for the correct annotation of genomic sequences and for the functional analysis of genes and their products. To elucidate the functions of a large population of Chinese cabbage (Brassica rapa) genes and to search efficiently for agriculturally useful genes, we have been taking advantage of the full-length cDNA Over-eXpresser (FOX) gene hunting system. With oligo dT column it purify the each mRNA from the flower organs, leaf and stem tissue. And about 120,000 cDNAs from the library were transformed into $\lambda$-pFLCIII-F vector. Of which 115,000 cDNAs from the library were transformed into T-DNA binary vector, pBigs for transformation study. We used normalized full-length cDNA and introduced each cDNA into Arabidopsis by in planta transformation. Full-length Chinese cabbage cDNAs were expressed independently under the CaMV 35S promoter in Arabidopsis. Selfed seeds were harvested from transgenic Arabidopsis. We had selected 2,500 transgenic plants by hygromycin antibiotic tolerant test, and obtained a number of transgenic mutants. Each transgenic Arabidopsis was investigated in morphological changes, fertility and leaf colour. As a result, 285 possible morphological mutants were identified. Introduced cDNA was isolated by PCR amplification of the genomic DNA from the transgenic mutants. Sequencing result and BLAST analysis showed that most of the introduced cDNA were complete cDNAs and functional genes. Also, we examined the effect of Bromelain on enhancing resistance to soft rot in transgenic Chinese cabbage 'Osome'. The bromelain gene identified from FOX hunting system was transformed into Chinese cabbage using Agrobacterium methods. Transformants were screened by PCR, then RT-PCR and real time PCR were performed to analyze gene expression of cysteine protease in the T1 and T2 generations. The anti-bacterial activity of bromelain was tested in Chinese cabbages infected with soft rot bacteria. The results showed that the over-expressed bromelain gene from pineapple conferred enhanced resistance to soft rot in Chinese cabbage.

• KIEE International Transactions on Electrophysics and Applications
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• v.4C no.4
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• pp.145-148
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• 2004

In this study, a DNA chip with a microelectrode array was fabricated using microfabrication technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5' end were immobilized on the gold electrodes by a DNA arrayer. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 and concentrated at the electrode surface through association with the formed hybrid. This suggested that this DNA chip could recognize the sequence specific genes.

• Proceedings of the KIEE Conference
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• 2004.07c
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• pp.2125-2127
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• 2004

In this paper, a DNA chip with a microelectrode array was fabricated using microfabrication technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5 end were immobilized on the gold electrodes by DNA arrayer. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.19
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• pp.8027-8034
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• 2014

Genetic instability contributes to the development and progression of gastric cancer, one of the leading causes of cancer death worldwide. Microsatellite instability (MSI) has been hypothesized to be involved in carcinogenesis, althgough its mechanisms and exact roles in gastric cancer remain largely unknown. Our aim was to identify associated clinicopathological characteristics and prognostic value of MSI in gastric cancer and precancerous lesions including gastritis, metaplasia, dysplasia, and adenoma. Because mitochondrial DNA has a different genetic system from nuclear DNA, the results of both nuclear MSI and mitochondrial MSI in gastric cancer were reviewed. This review provides evidence that genetic instability of nuclear and mitochondrial DNAs contributes to early stages of gastric carcinogenesis and suggests possible roles in predicting prognosis.

• Journal of the Korean Institute of Electrical and Electronic Material Engineers
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• v.19 no.10
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• pp.960-965
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• 2006

This research aims to develop the multiple channel electrochemical DNA chip using microfabrication technology. At first, we fabricated a high integration type DNA chip array by lithography technology. Several probe DNAs consisting of thiol group at their 5-end were immobilized on the gold electrodes. Then target DNAs were hybridized and reacted. Cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. Therefore, it is able to detect a plural genes electrochemically after immobilization of a plural probe DNA and hybridization of non-labeling target DNA on the electrodes simultaneously. It suggested that this DNA chip could recognize the sequence specific genes.

• Applied Microscopy
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• v.18 no.2
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• pp.167-176
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• 1988

Extrachromosomal circular DNA complexes from erythrocytes and splenocytes isolated from Carassius carassius were examined by mica-press-absorption method. The method was described that released small polydisperse circular DNA molecules in situ from the erythrocytes and the splenocytes and that allows selective observation of the small circular DNA complexes bound to cellular components. The released polydisperse circular DNA complexes were absorbed preferentially on mica in a divalent cation-free medium then processed for electron microscopy. Small circular DNAs showed a heterogeneous size distribution of $2{\sim}10{\mu}m$ with a mean contour length of $4.3{\mu}m$ for the circulating erythrocytes and that of $0.7{\sim}3.6{\mu}m$ with a mean contour of length $2.04{\mu}m$ for the splencytes. Cells contained $100{\sim}300$ copies and $300{\sim}700$ copies obtained from the erythrocytes and the splenocytes, repectively. Possible biological functional implications for size distribution of extrachromosomal circular DNAs are discussed.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2002.05a
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• pp.281-282
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• 2002

Out of 20 primers, 6 generated a total of 1,111 major and minor RAPD bands, producing approximately 4.2 average polymorphic bands per primer in shortnecked clam (Ruditapes philippinarum) population from Anmyeondo. The Bandsharing value altered from 0.15 to 0.74, with the average f 0.51, as calculated by bandsharing analysis. The RAPD profiles obtained with DNAs of two populations from Anmyeondo and Seocheon, respectively, were considerably different (0.20 and 0.51, respectively). The varying degrees of difference among populations amy also be of relevance to the restricted hybridization of wild bivalve. Besides gene mapping and breeding applications, PCR-RAPD systems could be very useful for the rapid certification and quality control of seed production and for every projects based on PCR amplification of specific bivalve DNA fragments.