• 제목/요약/키워드: DD-PCR

검색결과 83건 처리시간 0.034초

Linkage Disequilibrium of Dopamine D2 Receptor Gene in the Korean Population

  • Kang, Byung-Yong;Oh, Sang-Duk;Lee, Kang-Oh
    • Toxicological Research
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    • 제20권1호
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    • pp.49-53
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    • 2004
  • The genetic basis of hypertension is complex, and has been considered to be associated with the dopamine D2 receptor gene (DD2R). Because association studies using the candidate gene approach may provide important clues regarding the pathogenesis of hypertension and establish basis for further study, we performed the association study on the relationship between genetic polymorphisms in the DD2R gene and hypertension in Koreans. Eighty nine patients with hypertension and 86 age-matched subjects with normal blood pressure were enrolled. Genomic DNA was extracted from peripheral blood leukocytes. PCR-RFLP analysis was performed to detect the three polymorphic Taq I sites in the DD2R gene. There were no significant differences in genotype, allele and haplotype distributions of any polymorphisms in the DD2R gene between two groups, respectively (P>0.05), although significant linkage disequilibriums among these polymorphic sites were detected by pair-wise analysis (P<0.05). Therefore, our negative result suggest that the three Taq I RFLPs in the DD2R gene were not significantly associated with hypertension in Koreans.

DD-PCR을 이용한 벤조피렌 노출 조피볼락의 차등 발현 유전자 분석 (Analysis of Gene Expression in Benzo[a]pyrene-exposed Sebastes schlegeli using Differential Display Polymerase Chain Reaction)

  • 염승식;우선옥;최은석;김소정;오로라;이석찬;이택견
    • Environmental Analysis Health and Toxicology
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    • 제20권1호
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    • pp.67-73
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    • 2005
  • 오염물질의 노출에 의해 발현이 변화되는 유전자의 발굴은 외부환경 자극에 대한 적응이나 반응의 메커니즘을 알아내는뎨 중요한 정보를 제공하며, 오염물질에 반응하는 유전자는 환경오염을 감지하는 분자 마커로 개발될 수 있다. DD-PCR 기법은 차등 발현 유전자들을 발굴해내기 위한 유용한 방법으로 사용되어 왔고, 본 연구는 이 방법을 이용하여 벤조피렌에 반응하는 조피볼락 유전자들의 발굴을 목적으로 진행되었다. 간조직에서 추출한 RNA로부터 벤조피렌의 노출에 의해 발현 양이 달라진 12개의 클론을 발굴하였고, 그 염기서 열을 분석하였다. 또한 벤조피렌의 노출시간을 각각 6, 12, 24시간으로 달리한 조피볼락에서 12개의 클론 중 4개의 클론에 대해 northern blot 분석이 실시되었으며, 이들 모두 노출시간에 따 라 발현양이 증가 또는 감소하는 것이 확인되었다. 본 연구결과는 오염물질의 영향에 의한 유전자들의 발현에 관한 전반적인 지식을 제공하였고, 나아가 환경오염이나 외부 스트레스를 감지해 낼 수 있는 바이오마커의 개발을 위한 첫 단계로서의 정보를 제공할 수 있을 것으로 생각된다.

A novel WD40 protein, BnSWD1, is involved in salt stress in Brassica napus

  • Lee, Sang-Hun;Lee, Jun-Hee;Paek, Kyung-Hee;Kwon, Suk-Yoon;Cho, Hye-Sun;Kim, Shin-Je;Park, Jeong-Mee
    • Plant Biotechnology Reports
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    • 제4권2호
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    • pp.165-172
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    • 2010
  • Genes that are expressed early in specific response to high salinity conditions were isolated from rapeseed plant (Brassica napus L.) using an mRNA differential display method. Five PCR fragments (DD1.5) were isolated that were induced by, but showed different response kinetics to, 200 mM NaCl. Nucleotide sequence analysis and homology search revealed that the deduced amino sequences of three of the five cDNA fragments showed considerable similarity to those of ${\beta}$-mannosidase (DD1), tomato Pti-6 proteins (DD5), and the tobacco harpin-induced protein hin1 (DD4), respectively. In contrast, the remaining clones, DD3 and DD2, did not correspond to any substantial existing annotation. Using the DD3 fragment as a probe, we isolated a full-length cDNA clone from the cDNA library, which we termed BnSWD1 (Brassica napus salt responsive WD40 1). The predicted amino-acid sequence of BnSWD1 contains eight WD40 repeats and is conserved in all eukaryotes. Notably, the BnSWD1 gene is expressed at high levels under salt-stress conditions. Furthermore, we found that BnSWD1 was upregulated after treatment with abscisic acid, salicylic acid, and methyl jasmonate. Our study suggests that BnSWD1, which is a novel WD40 repeat-containing protein, has a function in salt-stress responses in plants, possibly via abscisic acid-dependent and/or -independent signaling pathways.

Identification of Novel Alternatively Spliced Transcripts of RBMS3 in Skeletal Muscle with Correlations to Insulin Action in vivo

  • Lee, Yong-Ho;Tokraks, Stephen;Nair, Saraswathy;Bogardus, Clifton;Permana, Paska A.
    • 대한의생명과학회지
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    • 제15권4호
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    • pp.301-307
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    • 2009
  • Whole-body insulin resistance results largely from impaired insulin-stimulated glucose disposal in skeletal muscle. Our previous studies using differential display and quantitative real-time RT-PCR have shown that a novel cDNA band (DD23) had a higher level of expression in insulin resistant skeletal muscle and it was correlated with whole-body insulin action, independent of age, sex, and percent body fat. In this study, we cloned and characterized DD23. The DD23 sequence is part of the 3'UTR region of the RNA binding motif, single stranded interacting protein (RBMS3). We have cloned the full length cDNA for RBMS3 and identified two splice variants. These variants named DD23-L and DD23-S have 15 and 14 exons respectively and differ from RBMS3 in the 3'UTR significantly. Northern blot analyses showed that an ~8.8 kb mRNA transcript of DD23 was predominantly expressed in skeletal muscle and to a lesser extent in placenta, but not in heart, brain, lung, liver, or kidney, unlike RBMS3. Elevated expression levels of these novel alternatively spliced variants of RBMS3 in skeletal muscle may play a role in whole body insulin resistance.

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한국인(韓國人) 구안와사(口眼喎斜) 환자(患者)의 Angiotensin Converting Enzyme 유전자(遺傳子) 다형성(多形成)에 관(關)한 임상연구(臨床硏究) (The Clinical Study on Angiotensin Converting Enzyme Gene Polymorphism in Korean Facial nerve palsy Patients)

  • 홍장무;박동석;고형균
    • Journal of Acupuncture Research
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    • 제21권2호
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    • pp.103-113
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    • 2004
  • Objective : This study was designed to investigate the relation between the angiotensin converting enzyme(ACE) gene polymorphism and Facial nerve palsy in the Korean population. Methods : This sudy was carried out on 117 Facial nerve palsy patients who were treated in the department of acupuncture & moxibustion, Hospital of Oriental medical college, Kyung-Hee University and 135 healthy control subjects. Blood samples from all subjects were obtaind for DNA extraction. The extracted DNA was amplified by polymerase chain reaction(PCR). PCR products were visualized by 2% agarose gel electrophoresis. Results : The sub-genotypes of ACE gene were II homozygotes, ID heterozygotes, DD homozygotes. While the distribution of ACE polymorphism in control subjects was 33%, 43%, 24%, the distribution of it in Facial nerve palsy patients was 32%, 50%, 18%(II, ID, DD). Thus, there was no significant different between the control and Facial nerve palsy groups. Conclusions : we conclude that there is no significant association between ACE gene polymorphism and Facial nerve palsy in Korean population. However, the findings of this study need to be confirmed in more patients and further studies. Additional epidemiologically based studies of the effects and relationship between ACE or other genes and lifestyles with regard to Facial nerve palsy is required.

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한국인(韓國人) 중풍(中風) 환자(患者)의 Angiotensin Converting Enzyme 유전자(遺傳子) 다형성(多形成)에 관(關)한 연구(硏究) (Angiotensin Converting Enzyme(ACE) Gene Polymorphism in Korean Stroke Patients)

  • 하지영;김창환;고형균
    • Journal of Acupuncture Research
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    • 제20권2호
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    • pp.161-172
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    • 2003
  • Objective : This study was designed to investigate the relation between the angiotensin converting enzyme(ACE) gene polymorphism and stroke in the Korean population. Methods : This study was carried out on 58 stroke patients who were hospitalized in the department of acupuncture & moxibustion, college of Oriental Medicine, Kyung-Hee University and 61 healthy control subjects. Blood samples from all subjects were obtaind for DNA extration. The extracted DNA was amplified by polymerase chain reaction(PCR). PCR products were visualized by 2% agarose gel electrophoresis. Results : The sub-genotypes of ACE gene were II homozygotes, ID heterozygotes, DD homozygotes. While the distribution of ACE polymorphism in control subjects was 31%, 51%, 18%, the distribution of it in stoke patients was 33%, 52%, 16%(II, ID, DD). Thus, there was no significant different between the control and stroke groups. Conclusions : we conclude that there is no significant association between ACE gene polymorphism and storke in Korean papulation. However, the findings of this study need to be confirmed in large patients and further studies. Additional epidemiologicallly based studies of the effects and relationship between ACE or other genes and lifesyles with regard to stroke required.

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Screening of Differentially Expressed Genes in Heterosigma akashiwo, a Red-Tide Causing Organism, Induced by Exposure to High Light

  • Ko, Young-Seok;Cho, Kyung-Je;Moon, Byoung-Yong
    • Journal of Photoscience
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    • 제8권3_4호
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    • pp.93-97
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    • 2001
  • Heterosigma akashiwo has been reported as red-tide causing phytoplankton in the Korean coastal area during summer when they are exposed to high light. It also shows photosynthetic adaptability to strong light during culture in the laboratory. On the basis of these observations, we tried to find out some genes specifically expressed in Heterosimga akashiwo during exposure to high light, assuming that they might have some resistant mechanisms associated with light adaptation. For this purpose, we carried out DD-PCR to detect differentially expressed mRNAs from cells that had been illuminated under high light for 3 days. We found eight cDNA clones that had been expressed specificically for high light. When they were further screened by reverse Northern hybridization, three of them were identified to be positive cDNA clones. When these cDNA fragments were subjected to DNA sequencing and then their base sequences were compared to GenBank database, one of them showed sequence homology 86% identical to the partial sequence of 16S rRNA gene of eubacterium CRO-18.

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Frequency of the Angiotensin - Converting Enzyme (ACE) Gene Polymorphism in the General Population and the Elite Endurance Students in Korea

  • Choung, Ho-Jin;Yoon, Song-Ro;Choi, Soo-Kyung
    • Journal of Genetic Medicine
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    • 제3권1호
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    • pp.11-13
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    • 1999
  • Recently it was reported that Insertion/Deletion polymorphism in the gene coding for Angiotensin-Converting Enzyme (ACE) is associated with human capacity for physical performance. This study was performed to genotyping of the ACE gene to determine the correlation between elite endurance performance and ACE I/D gene polymorphism. DNA sample was obtained from peripheral blood, hair roots and mouth epithelial cell in 739 general population and 200 elite athletic performance students. The ACE gene was amplified by polymerase chain reaction (PCR) using allele specific oligonucleotide primers. 155, 525 bp and 237 bp PCR products indicating the presence of insertion(I) and deletion(D) alleles, respectively, were clearly resolved after electrophoresis on a 2% agarose gel with ethidium bromide. Of the 200 elite athletic performance population subjects, 68(34%) showed ACE genotype 11,100(50%) genotype ID and 32(16%) genotype DD. Of the 739 general population subjects, 259(35.1%) showed ACE genotype 11,363(49.1%) genotype ID and 117(15.8%) genotype DD. Therefore ACE I/D gene polymorphism was not associated with human capacity for physical performance.(p>0.05)

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