• 제목/요약/키워드: PCR technique

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Multiple Symbiotic Associations Found in the Roots of Botrychium ternatum

  • Lee, Jun-Ki;Eom, Ahn-Heum;Lee, Sang-Sun
    • Mycobiology
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    • 제30권3호
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    • pp.146-153
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    • 2002
  • Two types of mycorrhizae, orchid(OM) and arbuscular mycorrhizae(AM), were observed in the cortical cells of Botrychium ternatum roots. The vesicles or arbuscules of AM fungi were examined and the fresh or digestive pelotons by other species of basidiomycetes were also observed in the roots under light microscope. These symbioses were, as the genomic DNAs extracted from roots of B. ternatum reacted with the specific primers, confirmed with PCR technique, being added to more strong evidences. These discoveries were rarely happened in the roots, especially a fern in nature. OM was observed in the roots of B. ternatum collected from the nationwide areas, whereas AM was only in the roots of B. ternatum collected from Chung-Buk areas. It is speculated that OM are associated with the nitrogen cycle in Islands and the growth of B. ternatum in the inland of Central Korea is related to both the phosphate and nitrogen cycle in the nature. The results suggest that B. ternatum is a typical species with two types of mycorrhizae under various growing conditions.

Involvement of D2 Receptor on Dopamine-induced Action in Interstitial Cells of Cajal from Mouse Colonic Intestine

  • Zuoa, Dong Chuan;Shahia, Pawan Kumar;Choia, Seok;Jun, Jae-Yeoul;Park, Jong-Seong
    • 대한의생명과학회지
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    • 제18권3호
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    • pp.218-226
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    • 2012
  • Dopamine is an enteric neurotransmitter that regulates gastrointestinal motility. This study was done to investigate whether dopamine modulates spontaneous pacemaker activity in cultured interstitial cells of Cajal (ICCs) from mouse using whole cell patch clamp technique, RT-PCR and live $Ca^{2+}$ imaging analysis. ICCs generate pacemaker inward currents at a holding potential of -70 mV and generate pacemaker potentials in current-clamp mode. Dopamine did not change the frequency and amplitude of pacemaker activity in small intestinal ICCs. On the contrary dopamine reduced the frequency and amplitude of pacemaker activity in large intestinal ICCs. RT-PCR analysis revealed that Dopamine2 and 4-receptors are expressed in c-Kit positive ICCs. Dopamine2 and 4 receptor agonists inhibited pacemaker activity in large intestinal ICCs mimicked those of dopamine. Domperidone, dopamine2 receptor antagonist, increased the frequency of pacemaker activity of large intestinal ICCs. In $Ca^{2+}$-imaging, dopamine inhibited spontaneous intracellular $Ca^{2+}$ oscillations of ICCs. These results suggest that dopamine can regulate gastrointestinal motility through modulating pacemaker activity of large intestinal ICCs and dopamine effects on ICCs are mediated by dopamine2 receptor and intracellular $Ca^{2+}$ modulation.

Enhancement of the Virtual Metrology Performance for Plasma-assisted Processes by Using Plasma Information (PI) Parameters

  • Park, Seolhye;Lee, Juyoung;Jeong, Sangmin;Jang, Yunchang;Ryu, Sangwon;Roh, Hyun-Joon;Kim, Gon-Ho
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2015년도 제49회 하계 정기학술대회 초록집
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    • pp.132-132
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    • 2015
  • Virtual metrology (VM) model based on plasma information (PI) parameter for C4F8 plasma-assisted oxide etching processes is developed to predict and monitor the process results such as an etching rate with improved performance. To apply fault detection and classification (FDC) or advanced process control (APC) models on to the real mass production lines efficiently, high performance VM model is certainly required and principal component regression (PCR) is preferred technique for VM modeling despite this method requires many number of data set to obtain statistically guaranteed accuracy. In this study, as an effective method to include the 'good information' representing parameter into the VM model, PI parameters are introduced and applied for the etch rate prediction. By the adoption of PI parameters of b-, q-factors and surface passivation parameters as PCs into the PCR based VM model, information about the reactions in the plasma volume, surface, and sheath regions can be efficiently included into the VM model; thus, the performance of VM is secured even for insufficient data set provided cases. For mass production data of 350 wafers, developed PI based VM (PI-VM) model was satisfied required prediction accuracy of industry in C4F8 plasma-assisted oxide etching process.

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시아노박테리아의 세포외산물에 대한 연구 (Extracellular Products from Cyanobacteria)

  • 권종희;김기은
    • KSBB Journal
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    • 제23권5호
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    • pp.398-402
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    • 2008
  • Cyanobacteria havebeen identified as one of the most promising group producing novel biochemically active natural products. Cyanobacteria are a very old group of prokaryotic organisms that produce very diverse secondary metabolites, especially non-ribosomal peptide and polyketide structures. Though many useful natural products have been identified in cyanobacterial biomass, cyanobacteria produce also extracellular proteins related with NRPS/PKS. Detection of unknown secondary metabolites in medium was carried in the present study by a screening of 98 cyanobacterial strains. A degenerated PCR technique as molecular approaches was used for general screening of NRPS/PKS gene in cyanobacteria. A putative PKS gene was detected by DKF/DKR primer in 38 strains (38.8%) and PCR amplicons resulted from a presence of NRPS gene were showed by MTF2/MTR2 primer in 30 strains (30.6%) and by A3/A7 primer in 26 strains (26.5%). HPLC analysis for a detection of natural products was performed in extracts from medium in which cyanobacteria containing putative PKS or NRPS were cultivated. CBT57, CBT62, CBT590 and CBT632 strains were screened for a production of extracellular natural products. 5 pure substances were detected from medium of these cyanobacteria.

Toward Functional Genomics of Plant-Pathogen Interactions: Isolation and Analysis of Defense-related Genes of Rot Pepper Expressed During Resistance Against Pathogen

  • Park, Do-Il;Lee, Sang-Hyeob
    • The Plant Pathology Journal
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    • 제18권2호
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    • pp.63-67
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    • 2002
  • To understand plant-pathogen interactions, a complete set of hot pepper genes differentially expressed against pathogen attack was isolated. As an initial step, hundreds of differentially expressed cDNAS were isolated from hot pepper leaves showing non-host resistance against bacterial plant pathogens (Xanthomonas campestris pv. glycines and Pseudomonas syringae pv. syringae) using differential display reverse transcription polymerase chain reaction (DDDRT-PCR) technique. Reverse Northern and Northern blot analyses revealed that 50% of those genes were differentially expressed in pepper loaves during non-host resistance response. Among them, independent genes without redundancy were micro-arrayed for further analysis. Random EST sequence database were also generated from various CDNA libraries including pepper tissue specific libraries and leaves showing non-host hypersensitive response against X. campestris pv. glycines. As a primary stage, thousands of cDNA clones were sequenced and EST data were analyzed. These clones are being spotted on glass slide to study the expression profiling. Results of this study may further broaden knowledge on plant-pathogen interactions.

AFLP 방법을 이용한 담배 버어리종 특이 프라이머의 개발 (Identification of tobacco Burley species specific marker in several tobacco species by AFLP)

  • 이영기;정석훈;금완수;이정헌;이청호;이문수
    • 한국연초학회지
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    • 제28권2호
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    • pp.94-99
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    • 2006
  • AFLP(Amplified Fragment Length Polymorphism) analysis was conducted to cultivars of tobacco, Nicotiana tabacum in order to select the cultivar-specific markers. AFLP results using 12 primer sets revealed genetic diversity among 12 field grown tobacco cultivars. Polymorphic fragments amplified by PCR was purified and cloned to identify their nucleotide sequences. From the sequences of them, 40 primer sets were designed to select cultivar-specific markers. When genomic DNA isolated from tobacco were used as PCR template, a set of primers, BrSF/BrSR showed Burley-specific band patterns. The results indicate that AFLP technique used in this experiments is useful for identifying tobacco cultivars in a rapid manner.

Propagation of lymphocystis disease virus (LCDV) in the FIN cell line originated from olive flounder Paralichthys olivaceus fin

  • Hossain, Mosharrof;Kim, Wi-Sik;Jung, Sung-Ju;Oh, Myung-Joo
    • 한국어병학회지
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    • 제24권2호
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    • pp.47-51
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    • 2011
  • The present study demonstrated lymphocystis disease virus (LCDV) propagation through cytopathic effects (CPE) formation and LCDV detection in olive flounder fin (FFN) cells by polymerase chain reaction (PCR) and fluorescent antibody technique (FAT) methods. Tissue filtrates from the cluster cells produced CPE in FFN cells, which initially cells became enlarged and gradually underwent fusion en masse. Infectivity of culture grown LCDV using the FFN cells reached $10^{2.3}$ $TCID_{50}$/ml at 4 days post infection and the highest titer was measured $10^{6.5}$ $TCID_{50}$/ml at 12 days. The viral DNA was detected in the cell culture supernatants showing CPE and the CPE cells by PCR. Antigen specific strong fluorescence reacting with monoclonal antibody against the virus revealed the presence of viral antigen in the cytoplasm of infected FFN cells. These results suggest that the FFN cell line originated from the olive flounder has a susceptibility of the LCDV.

Involvement of Cytochrome c Oxidase Subunit I Gene during Neuronal Differentiation of PC12 Cells

  • Kang, Hyo-Jung;Chung, Jun-Mo;Lee, See-Woo
    • BMB Reports
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    • 제30권4호
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    • pp.285-291
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    • 1997
  • It is becoming increasingly evident that significant changes in gene expression occur during the course of neuronal differentiation. Thus, it should be possible to gain information about the biochemical events by identifying differentially expressed genes in neuronal differentiation The PC12 cell line is a useful model system to investigate the molecular mechanism underlying neuronal differentiation and has been used extensively for the study of the molecular events that underlie the biological actions of nerve growth factor (NGF). In this study, we report an application of the recently described mRNA differential display method to analyze differential gene expression during neuronal differentiation. Using this technique, we have identified several cDNA tags expressed differentially during neuronal differentiation. Interestingly, one of these clones was cytochrome c oxidase subunit I (COX I) gene. The differential expression of COX I gene was confirmed by Northern blot analysis as well as RT-PCR. Southern blot analysis of the genomic DNA of PC12 cells revealed that COX I is a single gene. Induction of the oxidative enzyme might reflect the energy requirement in neuronal differentiation.

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The Gene Expression Profile of Cyst Epithelial Cells in Autosomal Dominant Polycystic Kidney Disease Patients

  • Lee, Jae-Eun;Park, Min-Ha;Park, Jong-Hoon
    • BMB Reports
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    • 제37권5호
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    • pp.612-617
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    • 2004
  • Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder characterized by the formation of fluid-filled cysts in the kidney and progressive renal failure. Other manifestations of ADPKD include the formation of cysts in other organs (liver, pancreas, and spleen), hypertension, cardiac defects, and cerebral aneurysms. The loss of function of the polycystin -1 and -2 results in the formation of epithelium-lined cysts, a process that depends on initial epithelial proliferation. cDNA microarrays powerfully monitor gene expression and have led to the discoveries of pathways regulating complex biological processes. We undertook to profile the gene expression patterns of epithelial cells derived from the cysts of ADPKD patients using the cDNA microarray technique. Candidate genes that were differently expressed in cyst tissues were identified. 19 genes were up-regulated, and 6 down-regulated. Semi-quantitative RT-PCR results were consistent with the microarray findings. To distinguish between normal and epithelial cells, we used the hierarchical method. The results obtained may provide a molecular basis for understanding the biological meaning of cytogenesis.

Characteristics of Purinergic Receptor Expressed in Human Retinoblastoma Cells

  • ;공인덕
    • 대한의생명과학회지
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    • 제13권4호
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    • pp.333-339
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    • 2007
  • Recently, much attention has been paid to human retinoblastoma since it provide a good model system for studying mechanisms underlying cell growth, differentiation, proliferation, and apoptosis, and for developing cancer therapy. However, until now it is unclear whether purinergic receptors are involved in the calcium mobilization in the retinoblastoma cells. In this regard, we measured possible purinergic signaling in WERI-Rb-1 cells using $Ca^{2+}$ imaging technique and RT-PCR method. ATP-induced $[Ca^{2+}]_i$ transients was maintained to about $90.7{\pm}1.0%$ of the control (n=48) even in the absence of extracellular calcium. The ATP-induced intracellular calcium response was only attained to $10.4{\pm}1.8%$ (n=55) of peak amplitude of the control after preincubation of 1 ${\mu}MU-73122$, a PLC inhibitor, but it was not affected by 1 ${\mu}MU-73343$, a inactive form of U-73122. And also ATP-induced $[Ca^{2+}]_i$ rise was almost attenuated by 20 ${\mu}M$ 2-APB, a putative $IP_3$ receptor inhibitor. Two subtypes of $IP_3$ receptor $(IP_{3-1}R,\;IP_{3-2}R)$ were identified by a RT-PCR method. These findings suggest that purinergic stimuli can cause calcium mobilization via $PLC-IP_3$ pathway after the activation of P2Y receptors in the retinoblastoma cells, which may play important roles in cell proliferation, differentiation, growth, and cell death.

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