• Title/Summary/Keyword: Human DNA

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Characterization of microsatellite markers covering chromosome 1 in the Korean and Japanese populations (한국인과 일본인에서 1번 염색체에 부착되는 microsatellite marker의 특징)

  • Lee, You-Jin;Park, Soo-Byung
    • The korean journal of orthodontics
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    • v.34 no.6 s.107
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    • pp.537-543
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    • 2004
  • Microsatellit markers are considered to be very promising genetic markers for genetic linkage analysis. The majority of the markers are as informative as in Caucasians but there are significant ethnic differences in the genetic variations. In order to investigate the genetic variations in the Korean and Japanese populations and their ethnic differences, 51 microsatellite marker loci spanning the whole human chromosome 1 were arranged from a commercially available set (ABI PRISM Linkage Mapping Set-HD5, Applied Biosystems, Foster City, CA, USA), and then determined the allelic frequencies and heterozygosities for these marker loci in the 90 unrelated Korean subjects and 90 unrelated Japanese subjects. Of all 51 markers tested, significant differences were observed when microsatellite allele frequency pattern of Korean was compared with those of Caucasian, while this pattern was highly similar between Korean and Japanese populations. Our data indicate that an extensive verification of public microsatellite markers in a particular population study should be undertaken prior to their linkage studies. Moreover, this information should facilitate genetic linkage studies of various hereditary diseases, especially in the Koreans and Japanese.

Sex Determination used Sex Determining Region Y Gene on the Y-chromosome of Human Teeth (사람 치아 Y염색체상의 sex determining region Y(SRY)유전자를 이용한 성별감정)

  • Kim, Sei-Youn;Ahn, Jong-Mo;Ryu, Geun-Chun;Yoon, Chang-Lyuk
    • Journal of Oral Medicine and Pain
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    • v.24 no.3
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    • pp.325-333
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    • 1999
  • 최근 중합효소연쇄반응을 이용한 분자생물학적 유전자분석기술의 발달로 성염색체상의 유전좌위 증폭을 통한 성별감정이 활발히 이루어지고 있다. 그중 사람 Y염색체상에 존재하는 남성 고환의 형성을 유도하는 sex-determining region Y(SRY) gene이 규명되어 유전질환의 조기 발견이나 예방 및 태아의 성별판정 등에 응용되고 있다. 그러나, 치아는 외부 환경에 대한 저항성이 가장 높은 장기로 성별감정 등 법의치과학적 개인식별에 널리 이용되고 있음에도 불구하고, SRY 유전자를 이용하여 치아에서의 성별감정에 대한 연구는 시도된 바 없다. 따라서, 본 연구에서는 사람 치아에서 중합효소연쇄반응법을 이용한 SRY 유전자를 검출하여 성별판정에 용용하고자 하였다. 남녀 각각 20개 치아의 치수와 상아질에서 DNA를 추출하여 중합효소연쇄반응 을 시행하고 SRY 유전자를 검색한 결과, 남성에서는 치수 13개중 8개, 상아질 7개중 4개에서 SRY 유전자가 검출되었고, 여생에서는 검출되지 않았다. 이러한 결과는 중합효소연쇄반응법을 이용하여 사람 치아에서 SRY 유전자를 검색할 때, 남성판별에 유용하고 치아를 이용한 성별감정시 기존의 성별감정에 이용되고 있는 다른 유전자와 함께 SRY 유전자를 검색함으로써 성별감정의 신뢰도를 높힐 수 있을 것으로 사료된다.

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Ectopic EBP2 expression enhances cyclin E1 expression and induces chromosome instability in HEK293 stable clones

  • Lee, Ming-Cheng;Hsieh, Chang-Hsun;Wei, Shu-Chen;Shen, Shu-Chen;Chen, Chiung-Nien;Wu, Vin-Cent;Chuang, Li-Ying;Hsieh, Fon-Jou;Wu, C. H. Herbert;Tsai-Wu, Jyy-Jih
    • BMB Reports
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    • v.41 no.10
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    • pp.716-721
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    • 2008
  • To explore the effects of deregulated expression of the EBNA1 binding protein 2 (EBP2) on cell growth, we generated human HEK293 stable clones constitutively expressing an EBP2-EGFP fusion protein. We found both RNA and protein levels of cyclin E1, a dominant oncoprotein, were elevated in the EBP2- EGFP stable clones. These findings were confirmed by flow cytometry bivariate analysis of cyclin expression versus DNA content. Moreover, the increase in p21 expression and the specific phosphorylation at Ser1981 of ATM and Ser15 of p53 were also observed in these stable clones, and these observations may explain the failure to observe an increase in Cdk2 kinase activity. In addition, after one year of passage culture, the EBP2-EGFP stable clones tended to lose 4 to 5 chromosomes per cell when compared to that of control cells. All of these findings provide a possible link between deregulated expression of EBP2 and tumor development.

Biodegradation of Endocrine Disrupting Chemicals by Genetic Transformants of Phlebia tremellosa Using Manganese Peroxidase Gene from Trametes versicolor (구름버섯 망간 과산화효소를 도입한 아교버섯 형질전환체에 의한 내분비장애 물질의 생분해)

  • Kum, Hyun-Woo;Kim, Myung-Kil;Choi, Hyoung-T.
    • Korean Journal of Microbiology
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    • v.45 no.1
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    • pp.82-85
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    • 2009
  • Endocrine disrupting chemicals (EDCs) disturb animal hormonal system even at very low concentrations, and finally give harmful effects to human through the food web. A white rot fungus Phlebia tremellosa isolated in Korea, was reported to have good degrading activity against the endocrine disrupting phthalates. However, this fungus has very low manganese peroxidase (MnP) activity under various culture conditions while laccase and lignin peroxidase activities were high. We have isolated an MnP cDNA from Trametes versicolor which was involved in the degradation of EDCs, and constructed an MnP expression vector to use in the genetic transformation of P. tremellosa in order to get higher MnP producing strains. Many transformants had integrated expression vector in their chromosomal DNAs, and showed increased MnP activity. One of two transformants showed increased degradation of 4 EDCs (70${\sim}$88%) than the wild type (30${\sim}$45% degradation rates), and showed twice better removal of estrogenic activities generated by the EDCs than the wild type.

Comparison of Four Multi-herbal Formulas in Antioxidant and Anticancer Activities (네 가지 처방의 항산화 및 항암 효과 비교)

  • Kim, Hong Jae;Choi, Eun Ok;Kim, Min Young;Son, Da Hee;Jeong, Jin-Woo;Lee, Sang-hyup;Park, Cheol;Choi, Yung Hyun;Hong, Su Hyun
    • Herbal Formula Science
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    • v.24 no.4
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    • pp.271-282
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    • 2016
  • Objective : In this study, we compared the antioxidant and anticancer properties of four multi-herbal formulas which were recorded in 'Dongeuibogam': Gilgyung-tang (GGT), Mokdanpi-tang (MDPT), Samso-eum (SSE), Samchulbobi-tang (SCBBT). Methods : We checked antioxidant properties of four multi-herbal formula through total phenolic content, radical scavenging activities, protective effects on genomic DNA oxidation. To investigate anticancer effects, we conducted MTT assay and analyzed morphologic change in A549 non-small lung cancer cells. Results : Total phenolic contents of four multi-herbal formulas were in a rich order of MDPT > SSE > GGT > SCBBT. Especially, MDPT revealed the highest activity than others in all antioxidant experiments. Our results indicated that treatment of those multi-herbal formulas induced growth retardation in A549 cells and MDPT also showed the highest anticancer effect ($IC_{50}=1.374mg/ml$) among them. Conclusions : Our data suggested that MDPT would be a powerful ingredient for lung cancer treatment.

Virulence, Resistance Genes, and Transformation Amongst Environmental Isolates of Escherichia coli and Acinetobacter spp.

  • Doughari, Hamuel James;Ndakidemi, Patrick Alois;Human, Izanne Susan;Benade, Spinney
    • Journal of Microbiology and Biotechnology
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    • v.22 no.1
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    • pp.25-33
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    • 2012
  • The association of verotoxic E. coli and Acinetobacter spp. with various antibiotic-resistant, diarrhogenic, and nosocomial infections has been a cause for concern worldwide. E. coli and A. haemolyticus isolated on a number of selective media were screened for virulence factors, antibiotic resistance, and transformation of resistance genes. Out of 69 E. coli isolates obtained, 25 (35.23%), 14 (20.30%), and 28 (40.58%) were positive for Vtx1&2, Vtx1, and Vtx2, respectively, 49 (71.015%) for extendedspectrum beta-lactamases (ESBLs), 34 (49.28%) for serum resistance, 57 (82.61%) for cell surface hydrophobicity, 48 (69.57%) for gelatinase production, and 37 (53.62%) for hemolysin production. For the 14 A. haemolyticus isolates, only 2 (14.29%) in each case from all the samples investigated were positive for Vtx1, Vtx2 and Vtx1&2 respectively, 8 (57.14%) for ESBLs, 7 (50.00%) for serum resistance, 11 (78.57%) for cell surface hydrophobicity, 4 (28.57%) for gelatinase production, and 8 (57.14%) for hemolysin production. Although transformation occurred among the E. coli and Acinetobacter isolates (transformation frequency: $13.3{\times}10^{-7}-53.4^{-7}$), there was poor curing of the plasmid genes, a confirmation of the presence of stable antibiotic-resistant genes (DNA concentration between 42.7 and 123.8 ${\mu}g$) and intragenetic transfer of multidrug-resistant genes among the isolates. The isolates were potentially virulent and contained potentially transferable antibiotic resistance genes. Detection of virulence factors, antibiotic resistance genes, and transformation among these isolates is a very significant outcome that will influence approaches to proactive preventive and control measures and future investigations. However, continued surveillance for drug resistance among these bacteria and further investigation of the mechanism of action of their virulence factors are a necessity.

Cloning and Characterization of Ginsenoside Ra1-Hydrolyzing ${\beta}$-D-Xylosidase from Bifidobacterium breve K-110

  • Hyun, Yang-Jin;Kim, Bo-Mi;Kim, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.535-540
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    • 2012
  • ${\beta}$-D-Xylosidase (E.C. 3.2.1.37) from Bifidobacterium breve K-110, which hydrolyzes ginsenoside Ra1 to ginsenoside Rb2, was cloned and expressed in Escherichia coli. The ($His_6$)-tagged recombinant enzyme, designated as XlyBK-110, was efficiently purified using $Ni^{2+}$-affinity chromatography (109.9-fold, 84% yield). The molecular mass of XylBK-100 was found to be 55.7 kDa by SDS-PAGE. Its sequence revealed a 1,347 bp open reading frame (ORF) encoding a protein containing 448 amino acids, which showed 82% identity (DNA) to the previously reported glycosyl hydrolase family 30 of Bifidobacterium adolescentis ATCC 15703. The $K_m$ and $V_{max}$ values toward p-nitrophenyl-${\beta}$-D-xylopyranoside (pNPX) were 1.45mM and 10.75 ${\mu}mol/min/mg$, respectively. This enzyme had pH and temperature optima at 6.0 and $45^{\circ}C$, respectively. XylBK-110 acted to the greatest extent on xyloglucosyl kakkalide, followed by pNPX and ginsenoside Ra1, but did not act on p-nitrophenyl-${\alpha}$-L-arabinofuranoside, p-nitrophenyl-${\beta}$-D-glucopyranoside, or p-nitrophenyl-${\beta}$-D-fucopyranoside. In conclusion, this is the first report on the cloning and expression of ${\beta}$-D-xylosidase-hydrolyzing ginsenoside Ra1 and kakkalide from human intestinal microflora.

Molecular Cloning of Vps26a, Vps26b, Vps29, and Vps35 and Expression Analysis of Retromer Complex in Micro Pig

  • Kim, Ek-Yune;Kim, Young-Hyun;Ryu, Chung-Hun;Lee, Jae-Woong;Kim, Sang-Hyun;Lee, Sang-Rae;Kim, Myeong-Su;Kim, Wan-Jun;Lim, Jeong-Mook;Chang, Kyu-Tae
    • Reproductive and Developmental Biology
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    • v.32 no.1
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    • pp.65-70
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    • 2008
  • Members of the Vps (Vacuolar protein sorting) protein family involved in the formation of the retromer complex have been discovered in a variety of species such as yeast, mouse, and human. A mammalian retromer complex is composed of Vps26, Vps29, and Vps35 proteins and plays and important role in cation-independent mannose-6-phosphate receptor retrieval from the endosome to the trans-Golgi network. In this study, we have identified the full-length sequences of the retromer components of Vps26, Vps29, and Vps35 in micro pigs. The cDNA sequences of these retromer components have been determined and the result showed there is 99% homology among the component counterparts from mouse, micro pigs, and humans. In addition, the retromer complexes formed with hetero-components were found in the brain of micro pigs. Based on above results, we suggest mammalian Vps components are well conserved in micro pigs.

Identification and Functional Characterization of Novel Genetic Variations in the OCTN1 Promoter

  • Park, Hyo Jin;Choi, Ji Ha
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.2
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    • pp.169-175
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    • 2014
  • Human organic cation/carnitine transporter 1 (OCTN1) plays an important role in the transport of drugs and endogenous substances. It is known that a missense variant of OCTN1 is significantly associated with Crohn's disease susceptibility. This study was performed to identify genetic variants of the OCTN1 promoter in Korean individuals and to determine their functional effects. First, the promoter region of OCTN1 was directly sequenced using genomic DNA samples from 48 healthy Koreans. OCTN1 promoter activity was then measured using a luciferase reporter assay in HCT-116 cells. Seven variants of the OCTN1 promoter were identified, two of which were novel. There were also four major OCTN1 promoter haplotypes. Three haplotypes (H1, H3, and H4) showed decreased transcriptional activity, which was reduced by 22.9%, 23.0%, and 44.6%, respectively (p<0.001), compared with the reference haplotype (H2). Transcription factor binding site analyses and gel shift assays revealed that NF-Y could bind to the region containing g.-1875T>A, a variant present in H3, and that the binding affinity of NF-Y was higher for the g.-1875T allele than for the g.-1875A allele. NF-Y could also repress OCTN1 transcription. These data suggest that three OCTN1 promoter haplotypes could regulate OCTN1 transcription. To our knowledge, this is the first study to identify functional variants of the OCTN1 promoter.

Influence of Rubiae Radix Extract on the Mechanism of Apoptosis in HL-60 Cells (천초근 추출물이 HL-60 세포주의 세포자멸사 기전에 미치는 영향)

  • Choi, Ho-Seung;Park, Jin-Mo;Ju, Sung-Min;Kim, Sung-Hoon;Kim, Dae-Keun;Kim, Won-Sin;Jeon, Byung-Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.3
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    • pp.548-555
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    • 2008
  • Rubiae radix belonging to the family Rubiaceae have been used in traditional medicine to blood stasis and hemostasis. In this study, we reported that methanol extract of Rubiae radix (RRME) induced apoptotic cell death through MAPKs activation in human promylocytic leukemia (HL-60) cells. The cytotoxic activity of activity of RRME in HL-60 cells was increased in a dose-dependent manner. RRME was cytotoxic to HL-60 cells, with IC50 of $8{\mu}g/mL$. Treatment of RRME to HL-60 cells showed apoptotic bodies, and the fragmentation of DNA, suggesting that these cells underwent apoptosis. Caspase-3 activity and PARP cleavage were time-dependently increased the expression of Bcl-2 and Bax. And ratio of Bax/Bcl-2 protein expression. Activation of p38 and JNK were increased 6 hr after RRME treatment in HL-60 cells, but activation of ERK was reduced 24 hr after treatment. Taken together, these results suggest that RRME induces apoptotic cell death through activation of p38 and JNK in HL-60 cells.