• Parasites, Hosts and Diseases
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• 제33권4호
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• pp.357-364
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• 1995

이질아메바에 의한 장염 환자의 조직 또는 이질아메바를 실험적으로 감염시킨 동물의 조직 검사에서 호중구의 침윤이 특징적으로 관찰된다. 그러나 이와같은 호중구의 침윤을 설명할 수 있는 기전에 대한 연구는 매우 미흡하다. 따라서 본 연구자들은 아메바 감염 초기에 인체 대장상피 세포에서 interleukin-8(IL-8)이 유도되어 호중구 침윤과 같은 염증반응이 유발될 것이라는 가설을 설정하였다. 이를 위하여 인체 대장상피세포주인 HT-29에 이질아메바 영양형을 실험적으로 노출시킨 뒤 발현되는 IL-S mRNA를 역전사 중합효소법(reverse transcriptional polymerase chain reaction, RT-PCR)으로 검사함과 퐁시에 발현된 IL-8 mRNA를 인공적으로 합성시킨 표준 RNA와 RT-PCR법을 이용하여 정량하였다. 실험 결과 이질아메바 영양형에 노출된 30분 후 부터 IL-8 mRAN가 발현되기 시작하였다 그리고 그 발현 분자수는 노출 시간의 증가에 따라 계속 증가하여 3시간 대에는 $3.1{\;}{\times}{\;}10^7{\;}molecules/\mu\textrm{g}$ total RNA를 나타내었다. 동시에 IL-8 mRNA의 발현은 노출시킨 이질아메바 영양형의 수에 비례하였다. 즉 HT-29/아메바 영양형의 비율이 10:1인 경우 IL-8 mRNA의 발현 분자수는 $1.2{\;}{\times}{\;}10^7{\;}molecules/\mu\textrm{g}$ total RNA로 나타났다. 이와같은 IL-8 mRNA의 발현은 IL-8 단백질 분비로 이어짐을 ELISA 검사로 확인할 수 있었다. 한편 이질아메바 파쇄액(Iysate)도 대장상피세포군인 Caco-2에서 IL-8 mRNA발현을 유도하였다. 결론적으로 본 실험은 이질아메바 감염 초기에 대장상피세포로 부터 IL-8이 발현되며, 이에 의하여 염증반응이 촉발될 가능성이 있음을 시사해 준다.

• Journal of Microbiology and Biotechnology
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• 제15권3호
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• pp.595-602
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• 2005

A one-step real-time quantitative RT-PCR assay in combination with automated RNA extraction was evaluated for routine testing of HCV RNA in the laboratory. Specific primers and probes were developed to detect 302 bp on 5'-UTR of HCV RNA. The assay was able to quantitate a dynamic linear range of $10^7-10^1$ HCV RNA copies/reaction ($R^2=0.997$). The synthetic HCV RNA standard of $1.84{\pm}0.1\;(mean{\pm}SD)$ copies developed in this study corresponded to 1 international unit (IU) of WHO International Standard for HCV RNA (96/790 I). The detection limit of the assay was 3 RNA copies/reaction (81 IU/ml) in plasma samples. The assay was comparable to the Amplicor HCV Monitor (Monitor) assay with correlation coefficient r=0.985, but was more sensitive than the Monitor assay. The assay could be completed within 3 h from RNA extraction to detection and data analysis for up to 32 samples. It allowed rapid RNA extraction, detection, and quantitation of HCV RNA in plasma samples. The method provided sufficient sensitivity and reproducibility and proved to be fast and labor-saving, so that it was suitable for high throughput HCV RNA test.

• 미생물학회지
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• 제2권1호
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• pp.1-11
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• 1964

Yung Nok Lee (Dept. of Biology, Korea University) : Studies on the phosphate metabolism in Chlorella, with special reference to polyphosphate. Kor. J. Microbiol., Vol.2, No.1, p1-11 (1964). 1. Uniformly $^{32}P$-labeled Chlorella cells which were irradiated with Cobalt-60 gamma-rays of about 70, 000 $\gamma$ dose, were further grown in a standard "cold" medium ("hot".rarw."cold"), and some portions of the algae were taken out at the begining of, and at intervals during the culture, and subjected to analyze the contents of $^{32}P$- and total P in various fractions of the cell materials. Results obtained were compared with those of nonirradiated normal cells. 2. Amounts of phosphate in various fractions of the nonirradiated normal Chlorella cells were measured using uniformly $^{32}P$--labeled cells. Analysis of the $^{32}P$--labeled algal cells showed that the highest value in P-content was the fraction of RNA followed by those of lipid, polyphosphate "C" polyphosphate "B", DNA, nucleotidic labile phosphate compounds, polyphosphate "A" and protein. It was observed that content of total polyphosphates in a single Chlorella cell was almost equal to RNA-P content in the cell, and the amount of RNA-P was almost equal to ten times of DNA-P content. 3. When the $^{32}P$--labeled algae which were irradiated with gamma-rays were grown in a normal "cold" medium, phosphate contents in the fraction of DNA, nucleotidic labile phosphate compounds and protein decreased markedly, while the contents of phosphate in the fractions of polyphosphate "C" and potyphosphate "B" increased in comparison with those of unirradiated normal cells. So, it was considered that the pretreatment of above mentioned dose of gamma-ray inhibited DNA and protein synthesis from polyphosphate in Chlorella cells. 4. Proceeding the culture of $^{32}P$--labeled Chlorella in a "cold" standard medium, whose synthetic activity of DNA and protein from polyphosphate was disturded by gamma-ray irradiation, the amounts of $^{32}P$-in the fraction of polyphosphate "C" increased, in contrast with those of polyphosphate "B" fraction. According to these experimental results, it was inferred that polyphosphate "B" could transform into polyphosphate "C" in normal growing Chlorella cells.sults, it was inferred that polyphosphate "B" could transform into polyphosphate "C" in normal growing Chlorella cells.ing Chlorella cells.

• BMB Reports
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• 제29권2호
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• pp.122-126
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• 1996

A series of oligonucleotides were synthesized by automatic DNA synthesizer. The purity of crude products was checked and their molecular weights determined by matrix assisted laser desorption ionization mass spectrometry (MALDI-MS) with an accuracy of better than 0.05% deviation even without using an internal standard. This mass determining technology in combination with partial digestion of oligonucleotides by 5'- and 3'-exonuclease provides a straightforward and simple method to obtain sequence information of oligonucleotides. The extension of this technology to the sequencing of modified oligonucleotides and genomic DNA and RNA might become possible.

• 생명과학회지
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• 제34권8호
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• pp.588-593
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• 2024

대사경로 3,490개가 고세균 471종에 분포하는 정도를 MetaCyc database의 자료를 이용하여 분석하였다. 대사경로의 수는 고세균의 종류에 따라 13~184개가 존재하였다. 모든 고세균에 공통적으로 존재하는 대사경로는 없었으며 470종의 고세균에 UTP and CTP de novo biosynthesis와 tRNA charging의 대사경로가 존재하였다. 고세균들에 분포하는 상위 12개의 대사경로 중에서 핵산관련이 5개, 단백질관련이 5개 그리고 생체의 여러 반응에 참여하는 cofactor인 S-adenosyl-L-methionine (SAM)의 생합성 대사경로와 효소의 활성에 필수적인 번역 후 변형에 필요한 phosphopantothenate biosynthesis III (archaea)의 대사경로로 나타나 핵산과 물질대사와 관련된 단백질의 중요성을 알 수 있었다. 대사경로 보유 계통수에서 구한 distance value를 이용하여 고세균의 각 강(class)을 평균과 표준편차로 나누었을 때, 2개의 그룹과 기타로 나뉘어져 대사경로의 분포가 다양한 것으로 나타났다. 본 연구결과는 기초과학 이외에 약물개발 등에도 응용될 수 있을 것으로 기대된다.