• Title/Summary/Keyword: DNA technology

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이달의 과학자 - 포항공대 생명과학과 교수 '성영철'

  • Korean Federation of Science and Technology Societies
    • The Science & Technology
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    • v.33 no.2 s.369
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    • pp.18-19
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    • 2000
  • 포항공대 성영철 교수는 최근 에이즈의 DNA백신을 개발, 원숭이에서 방어면역이 발생함을 증명해내 에이즈 완치에 청신호가 되고 있다. 세계 최초로 개발에 성공한 이 이에즈 DNA백신은 미국 식품의약국(FDA)에서도 안전하다고 인정하고 있는 순수 DNA만으로 제조된 것이어서 사람에 대한 임상실험이 신속히 이루어질 수 있는 것이다. 이번 연구결과는 이미 특허출원했으며 소동물(생쥐) 실험결과는 백신분야의 세계적 권위지 "vaccine"지에 발표되었다.

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Construction of Human cDNA Library Analysis Pipeline (인간 cDNA 라이브러리 분석 파이프라인 구축)

  • Jung, Jaeeun;Kim, Dae-Soo
    • Proceedings of the Korea Contents Association Conference
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    • 2018.05a
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    • pp.323-324
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    • 2018
  • 전장 cDNA 클론을 시퀀싱하는 것은 선택적 스플라이싱 형태를 비롯한 정확한 유전자 구조를 정의하는데 유용하게 사용될 수 있으며, 유전자 및 단백질의 생물학적 기능연구에 중요한 자원을 제공한다. 포괄적이며 비 중복적인 cDNA의 생산은 인간 유전체 연구의 중요한 목표이다. 본 연구에서 제공하는 인간 cDNA 라이브러리 분석 파이프라인은 전장 cDNA를 분석하는 자동화 도구로 여러 연구자들에게 활용 될 수 있을 것으로 사료된다.

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Construction of Human cDNA Library Analysis Pipeline (인간 cDNA 라이브러리 분석 파이프라인 구축)

  • Jung, Jaeeun;Kim, Dae-Soo
    • Proceedings of the Korea Contents Association Conference
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    • 2018.05a
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    • pp.83-84
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    • 2018
  • 전장 cDNA 클론을 시퀀싱하는 것은 선택적 스플라이싱 형태를 비롯한 정확한 유전자 구조를 정의하는데 유용하게 사용될 수 있으며, 유전자 및 단백질의 생물학적 기능연구에 중요한 자원을 제공한다. 포괄적이며 비 중복적인 cDNA의 생산은 인간 유전체 연구의 중요한 목표이다. 본 연구에서 제공하는 인간 cDNA 라이브러리 분석 파이프라인은 전장 cDNA를 분석하는 자동화 도구로 여러 연구자들에게 활용 될 수 있을 것으로 사료된다.

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Toxicogenomic analysis of Effects of Bisphenol A on Japanese Medaka fish using high density-functional cDNA microarray

  • Jiho Min;Park, Kyeong-Seo;Hong, Han-Na;Gu, Man-Bock
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.173-173
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    • 2003
  • With the introduction of DNA microarrays, a high throughput analysis of gene expression is now possible as a replacement to the traditional time-consuming Southern-blot analysis. This cDNA microarray should be ahighly favored technology in the area of molecular toxicology or analysis of environmental stresses.In this study, therefore, we developed a novel cDNA microarray for analyzing stress-specific responses in japanese Medaka fish. In the design and fabrication of this stress specific functional cDNA microarray, 123 different genes in Medaka fish were selected from eighteen different stress responsive groups and spotted on a 25${\times}$75 mm glass surface. After exposure of the fish to bisphenol A which is the one of the well-known endocrine disrupting chemicals (EDCs), over 1 or 10 days, the responses of the DNA chip were found to show distinct expression patterns according to the mode of toxic actions from environmental toxicants. As a results, they showed specific gene expression pattern to bisphenol A, additionally, the chemical spesific biomarkers could be suggested based on the chip analysis data. Therefore, this chip can be used to monitor stress responses of unknown and/or known toxic chemicals using Medaka fish and may be used for the further development of biomarkers by utilizing the gene expression patterns for known contaminants.

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Cloning of An Intron of the Gene Coding for Porcine Acid-Labile Subunit(pALS) of the 150-kDa Insulin-like Growth Factor Complex and the 3' ntranslated Region of pALS Complementary DNA and Confirmation of pALS Gene Expression in Multiple Tissues (돼지 150-kDa Insulin-like Growth Factor Complex의 Acid-labile Subunit(ALS) 유전자의 Intron 및 ALS Complementary DNA의 3' 비해독 부위 Cloning과 생체조직에서의 ALS 유전자 발현 확인)

  • Jin, E.J.;Kim, I.A.;Lee, C.Y.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.555-562
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    • 2004
  • 본 연구는 목저은 다음과 같다: 1) 돼지에서 150-kDa temary insulin-like growth faetor(IGF)complex의 한 구성 요소인 acid-labile subunit(ALS) 유전자 intron의 존재 확인. cloning 및 돼지 ALS(porcine ALS; pALS) complementary DNA(cDNA)의 3' 비해독(untranslated) 부위(3' UT) 증폭. cloning, 2) intron-spanning primer pair를 이용한 reverse transcription-polymerase chain reaction(RT-PCR) 방법에 의한 돼지 조직에서의 ALS 유전자 발현 분포 확인 및 3) 돼지 hepatocyte에서의 ALS 유전자 발현 여부 확인. 돼지 genomic DNA를 template로 하여 PCR 방법으로 예상된는 intron 부위를 증폭하고 plasmid vector에 삽입하여 염기서열을 결정한 결과 타 종의 ALS 유전자에서와 같은 위치에 1,371-base pair(bp)의 pALS intron이 존재함을 확인하였다. 역시 본 연구에서 간에서 추출한 RNA를 주형으로 시작하여 3' rapid amplification of cDNA end(3' RACE) 방법으로 147-bp의 3'UT를 합성하고 그 염기성열을 결정하였다. RT-PCR 결과 간은 물론 조사된 모든 돼지의 내장기관(신장, 폐, 비장)과 자성 생식기관(난소, 난관, 자궁) 및 골격근육에서 ALS 유전자가 발현됨이 밝혀졌다. 또한 돼지 간 조직에 대한 in-situ hybridization 결과 hepatocyte에서 ALS 유전자가 발현됨이 확인되었다. 이상의 결과는 ALS가 혈중 IGF의 저정/조절체로서의 주기능 외에 모세혈관 밖에서도 미지의 기능이 있을 기능성을 시사한다.

Production of DNA polymerase from Thermus aquaticus in recombinant Escherichia coli

  • Kim, Sung-Gun;Park, Jong-Tae
    • Korean Journal of Agricultural Science
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    • v.41 no.3
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    • pp.245-249
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    • 2014
  • Among dozens of DNA polymerases cloned from thermophilic bacteria, Taq DNA polymerase from Thermus aquaticus has been most frequently used in polymerase chain reaction (PCR) that is being applied to gene cloning, DNA sequencing, gene expression analysis, and detection of infectious and genetic diseases. Since native Taq DNA polymerase is expressed at low level in T. aquaticus, recombinant Escherichia coli system was used to produce Taq DNA polymerase in a large amount. Taq DNA polymerase was expressed as a soluble form under the control of tac promoter in E. coli, and purified by heat treatment and ion exchange chromatographies. The purified Taq DNA polymerase was nearly homogeneous and exhibited a similar DNA amplification activity with a commercial Taq DNA polymerase.

Full Length cDNA, Genomic Organizations and Expression Profiles of the Porcine Proteasomal ATPases PSMC5 Gene

  • Wang, Y.F.;Yu, M.;Liu, B.;Fan, B.;Wang, H.;Zhu, M.J.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.7
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    • pp.897-902
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    • 2004
  • PSMC5 subunit, which belongs to the 26S proteasomal subunit family, plays an important role in the antigen presentation mediated by MHC class I molecular. Full-length cDNA of porcine PSMC5 was isolated using the in silico cloning and rapid amplification of cDNA ends (RACE). Amino acid was deduced and the primary structure was analyzed. Results revealed that the porcine PSMC5 gene shares the high degree of sequence similarity with its mammalian counterparts at both the nucleotide level and the amino acid level. The RT-PCR was performed to detect the porcine PSMC5 expression pattern in seven tissues and the result showed that high express level was observed in spleen, lung, marrow and liver while the low express level was in muscle. The full-length genomic DNA sequence of porcine PSMC5 gene was amplified by PCR and the genomic structure revealed that this gene was comprised by 12 exons and 11 introns. Best alignment of the cDNA and genomic exon DNA sequence presents 4 mismatches and this information potentially bears further study in gene polymorphisms.

Comparison of methods of DNA extraction from tree nuts (견과류로부터 효율적인 DNA 추출 방법 비교)

  • Suh, Seung-Man;Park, Saet-Byul;Kim, Mi-Ju;Kim, Hae-Yeong
    • Korean Journal of Food Science and Technology
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    • v.50 no.4
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    • pp.357-361
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    • 2018
  • This study aimed to explore efficient DNA extraction methods using tree nuts. Four different DNA extraction procedures, including silica membrane method, modified silica method, cetyltrimethylammonium bromide (CTAB) method, and modified CTAB method were examined for their relative efficiency in extracting DNA from pistachio, pine nut, almond, hazelnut, cashew nut, walnut, and peanut. The quality and quantity of the extracted DNA were subsequently assessed by spectrometric measurements, gel electrophoresis, and PCR amplifications. CTAB method was the most appropriate one for extracting DNA from pine nut, cashew nut, pistachio, and peanut. However, it could be replaced by the silica membrane method for walnut and modified CTAB method for almond and hazelnut.

The Specific Probes Confirming the Genomic DNA of Tricholoma matsutake in Korea (송이의 Genomic DNA에 특이적인 Probe)

  • Lee, Sang-Sun;Hong, Sung-Woon;Chung, Hung-Chae;Sung, Chang-Kun;Kim, Jae-Hun;Ka, Kang-Hyeon;Kim, Hyun-Joong
    • The Korean Journal of Mycology
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    • v.27 no.1 s.88
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    • pp.20-26
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    • 1999
  • The specific DNA band appeared in PCR-RAPD analysis using OPO-2 primer was a very important for the researching Korean pine-mushrooms, Tricholoma matsutake. This DNA band, sequenced to be the 770 base pairs, existed as only a single copy in the whole genomic DNA's of Korean pine-mushrooms. However, this band was not presenting from the PCR-RAPD bands of other ectomycorrhyzal fungi reacted with the OPO-2 primer or the dot blots. Also, this DNA sequence was not matched with those of the other genes known by NCBI and had low homology together with sequence of other proteins compared. Those results suggested that the specific DNA band can be used as probe for identification of T. matsutake and might be related to the informations rather than the gene for the proteins with analysis of protein sequence translated from the DNA sequence.

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Desmutagenicity of Enzymatically Browned Substances Obtained from the Reaction of Prunus salicina (Red) Enzyme and Polyphenols (재래종 적색자두(Prunus salicina) 효소갈변반응 생성물의 돌연변이 억제작용)

  • Ham, Seung-Shi;Hong, Eun-Hee;Omura, Hirohisa
    • Korean Journal of Food Science and Technology
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    • v.19 no.3
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    • pp.212-219
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    • 1987
  • The rec-assay on Bacillus subtilis strains H17$({Rec}^+)$ and M45$({Rec}^-)$, the Ames test with modification of preincubation on Salmonella typhimurium TA98 and TA100 and DNA-breaking test on double strand calfthymus DNA were carried out using enzymatically browned substances obtained from the reaction of Prunus salicina (Red) enzyme and polyphenols. The spore rec-assay of enzymatic browning reaction products of pyrogallol, hydroxyhydroquinone. 3,4-dihydrohyoluene and chlorogenic acid showed non-mutagenic activity The spore rec-assay showed a little influence of ${Zn}^{2+}$ and ${Ni}^{2+}$ on the action of four kinds of enzymatic browning reaction products. The enzymatic browning reaction products of polyphenols did not show DNAbreaking activity. ${Cu}^{2+}$ of various metal ions influenced on DNA-breaking of enzymatic browning reaction products of pyrogallol. However, enzymatic browning reaction products of chlorogenic acid inhibited on DNA-breaking activity. Four kinds of enzymatic browning reaction products showed non-mutagenic activity on Salmonella typhimurium TA98 and TA100 with S-9 mix. In the mutagenicity on Salmonella typhimurium TA98 and TA100 with S-9 mix in the presence of benzo$({\alpha})$pyrene which is the carcinogenic substances, four kinds of enzymatic browning reaction products showed desmutagenic activity.

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