• 제목/요약/키워드: Xa1

검색결과 71건 처리시간 0.02초

남성과 여성에서 XIST 유전자의 후성학적 비교 연구 (Epigenetic Study of XIST Gene from Female and Male Cells by Pyrosequencing)

  • 김환희;윤여진;송민애;이수만
    • Clinical and Experimental Reproductive Medicine
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    • 제37권1호
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    • pp.25-31
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    • 2010
  • 목 적: X 염색체 불활성화는 여성과 남성 사이에 X 염색체의 유전자 발현 유지를 위해 여성의 X 염색체 중 하나가 불활성화 되는 현상이다. 이러한 X 염색체 불활성화는 해독되지 않는 XIST 유전자에 의해 조절된다. XIST 유전자는 오직 불활성화된 X 염색체 에서만 발현되고, 활성화된 X 염색체 에서는 발현되지 않는다. 따라서 체세포에서 활성화된 X 염색체의 XIST 유전자는 promoter 부분이 메틸화 되어있고, 불활성화된 X 염색체에서는 메틸화가 거의 되어 있지 않다. 연구방법: 본 연구에서는 정상 여성과 정상 남성의 XIST 유전자의 promoter와 5'-end 지역의 메틸화 차이를 측정하기 위해 정상여성과 남성의 혈액에서 DNA를 추출하여 파이로시퀀싱 (Pyrosequencing) 방법을 통해 XIST 유전자의 총 8부분의 CpG 영역 (-1696, -1679, -1475, -1473, -1469, +947, +956, +971)을 분석하였다. 결 과: 총 8부분의 CpG 영역을 분석한 결과, promoter 부분인 CpG 1-5 영역 (-1696, -1679, -1475, -1473, -1469)에서는 여성과 남성의 메틸화 정도에 차이가 없었다. 그러나 5'-end 부분인 CpG6-8 영역 (+947, +956, +971)에서는 여성이 45.2% 49.9% 44.2%, 남성이 90.6%, 96.7%, 87.8%으로 메틸화 정도가 차이를 나타냈다. 결 론: 따라서 본 연구에 사용한 방법은 XIST 유전자의 메틸화 패턴의 차이를 기존의 방법보다 신속하고 정확하게 분석할 수 있다는 장점이 있기 때문에 유용하게 사용될 수 있을 것이다.

누에 배양세포(Bm5) 및 생체에서 베큘로바이러스 발현계를 이용한 누에신 단백질 발현 특성 (Expression of Antibacterial Protein, Nuecin, Using Baculorivus Expression Vector System in Bm5 Insect Cell and Bombyx mori)

  • 윤은영;구태원;황재삼;김상현;강석우;김근영;진병래
    • 한국잠사곤충학회지
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    • 제44권2호
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    • pp.69-73
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    • 2002
  • 본 연구는 누에신 단백질의 대량발현을 통해 농업용 소재로서 이용하고자 하는 연구의 일환으로 누에 핵다각체병 바이러스 유래의 pBm10po1-Xa 벡터에 누에신 유전자를 도입하여 누에 배양세포(Bm5) 및 누에 생체에서 발현한 결과 누에신 전사체는 바이러스 접종 후 1일째부터 발현되기 시작하여 5일째에 최대로 발현되었음을 확인할 수 있었고, 누에신 단백질은 3일째부터 발현량이 증가하여 5일째까지 계속 지속적으로 발현되었으나 그 발현량은 전사체에서 처럼 많지 않음을 확인할 수 있었으며, 누에신 단백질의 발현량은 누에 세포에 비해 누에 생체에서 기대만큼 그 발현량이 많지 않았다. 또한 누에신의 베큘로바이러스 발현계 (baculovirus expression vector system, BEVS)를 이용하여 세포내 및 번역후 변형과정을 통하여 세포외로 분비된 누에신의 발현양상을 확인한 결과 세포내에 비해 세포외로 분비시 그 발현량이 현저히 줄어들었음을 확인할 수 있었다. 따라서 베큘로바이러스 발현계를 이용하여 외래 단백질을 생산할 경우 정확한 메카니즘은 밝혀지지 않았으나 강력한 프로모터에 의해 세포내 단백질 생산량은 많은데 비해 정확한 단백질 고차구조 형성을 도와주는 foldase 및 chaperon의 양은 한정되어져 있으므로 정확한 고차구조를 형성하여 세포외로 분비되는 생물학적 활성을 띠는 단백질은 매우 적은 양간이 발현됨을 확인할 수 있었다. 그러므로 추후 누에 신 단백질 대량생산을 위해서는 분비 프로세싱의 해명 및 번역 후 변형과정의 개선을 통한 발현계의 개량이 시급히 요구된다.

Characterization of the Pediocin Operon of Pediococcus acidilactici K10 and Expression of His-Tagged Recombinant Pediocin PA-1 in Escherichia coli

  • MOON GI SEONG;PYUN YU RYANG;KIM WANG JUNE
    • Journal of Microbiology and Biotechnology
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    • 제15권2호
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    • pp.403-411
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    • 2005
  • The relationship between plasmid (~9.5 kb) and pediocin PA-1 in P. acidilactici K10 was confirmed by plasmid curing. The pediocin operon of P. acidilactici K10 was amplified by PCR (polymerase chain reaction), and the nucleotide sequence was analyzed. The sequence of the pediocin operon of P. acidilactici K10 was similar to those of P. acidilactici strains producing pediocin PA-1/ AcH. For the expression of pediocin PA-1 in E. coli, a pQEPED (pQE-30 Xa::mature pedA) was constructed. His-tagged recombinant pediocin PA-1 (-6.5 kDa) was translated by cell-free in vitro transcription and translation using pQEPED as a DNA template. Theresult of slot blotting assay showed that transcription of recombinant pedA in E. coli M15 was induced by the addition of isopropyl-$\beta$-D-thiogalactopyranoside (IPTG) at the final concentration of 1 mM. Although the recombinant pediocin PA-1 inhibited the growth of E. coli, it was expressed in the host strain and purified by nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography under denaturing condition. This is the first report for the production and one-step purification of biologically active recombinant pediocin PA-1 in E. coli.

메밀 발아 중 식이섬유 함량과 조성의 변화 (Changes in Contents and Composition of Dietary Fiber during Buckwheat Germination)

  • 이명헌;우순자
    • 한국식품영양학회지
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    • 제7권4호
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    • pp.274-283
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    • 1994
  • To provide the efficient application scheme of buckwheat dietary fiber and basic information of seed germination, buckwheat(Fagopyrum esculentum Moench) was germinated at 10$^{\circ}C$ for 7 days and the contents and composition of the total dietary fiber(TDF), insoluble dietary fiber(IDF), soluble dietary fiber(SDF) wire examined at 24 hour intervals. The TDF content in ungerminated seeds was 24.86o on dry weight basis. It decreased for the 1st day of germination, but gradually increased for 7 days afterwords. The contents of IDF and SDF in ungerminated seeds were 22.05, 1.42% respectively. The IDF and SDF contents decreased in the initial stage of germination, but then gradually increased. The composition of the IDF and SDF in the TDF during the germination period showed different tendencies. The IDF decreased with germination time until 5 days and then increased. The SDV increased until 5 days and then decreased gradually. The TDF contents obtained by AOAC method were generally higher than those obtained by Prosky method. The TDF contents obtained by the two method, however, were very closely correlated (r=0.9966, p< 0.01) The IDF(X1) and SDF(X2) showed the significant regression equation(p<0.01) with the root length(Y). The equation was Y: -12.6681+0.5089${\times}$ 1 $\div$ 0.6022Xa and R2 was 0.968.

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Deciphering the Role of Tyrosine Sulfation in Xanthomonas oryzae pv. oryzae Using Shotgun Proteomic Analysis

  • Park, Hye-Jee;Park, Chang-Jin;Bae, Nahee;Han, Sang-Wook
    • The Plant Pathology Journal
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    • 제32권3호
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    • pp.266-272
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    • 2016
  • A bacterial tyrosine sulfotransferase, RaxST, is required for activation of rice XA21-mediated immunity, and it catalyzes sulfation of tyrosine residues of Omp1X and RaxX in Xanthomonas oryzae pv. oryzae, a causal agent of bacterial blight in rice. Although RaxST is biochemically well-characterized, biological functions of tyrosine sulfation have not been fully elucidated. We compared protein expression patterns between the wildtype and a raxST knockout mutant using shotgun proteomic analysis. Forty nine proteins displayed a more than 1.5-fold difference in their expression between the wildtype and the mutant strains. Clusters of orthologous groups analysis revealed that proteins involved in cell motility were most abundant, and phenotypic observation also showed that the twitching motility of the mutant was dramatically changed. These results indicate that tyrosine sulfation by RaxST is essential for Xoo movement, and they provide new insights into the biological roles of RaxST in cellular processes.

Construction of a Novel Recombinant Bombyx mori Nuclear Polyhedrosis Virus Producing the Fluorescent Polyhedra

  • Kang, Seok-Woo;Yun, Eun-Young;Woo, Soo-Dong;Goo, Tae-Won;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • 제3권1호
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    • pp.75-81
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    • 2001
  • We have constructed a novel recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV) producing the green fluorescent polyhedra. For the production of the fluorescent polyhedra, partial polyhedrin gene containing KRKK as nuclear localization site from the BmNPV polyhedrin gene and the green fluorescent protein (gfp) gene were introduced under the control of p10 promoter of BmNPV. The recombinant BmNPV was stably produced fluorescent polyhedra in the infected Bm5 cells and the morphology of the fluorescent polyhedra was similar to that of wild-type BmNPV. The fluorescent polyhedra had 32 kDa native polyhedrin and 41 kDa fusion protein. From these data, we have further developed a novel BmNPV p10-based transfer vector producing recombinant polyhedra with foreign gene Product. The novel BmNPV P10-based transfer vector is composed of partial polyhedrin gene, factor Xa, and multiple cloning sites.

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Expression of an Antimicrobial Peptide Magainin by a Promoter Inversion System

  • Lee, Jae-Hyun;Hong, Seung-Suh;Kim, Sun-Chang
    • Journal of Microbiology and Biotechnology
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    • 제8권1호
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    • pp.34-41
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    • 1998
  • A method was developed for the controlled expression of an antimicrobial peptide magainin in Escherichia coli. A series of concatemeric magainin genes was constructed with a gene amplification vector, and fused to the 3'end of malE gene encoding the affinity ligand, E. coli maltose-binding protein (MBP). The construct directed the synthesis of the fusion protein with the magainin polypeptide fused to the C-terminus of MBP. The fusion protein was expressed in a tightly regulatable expression system which was under the control of an invertible promoter. The MBP-fused magainin monomer was expressed efficiently. However, the expression level of the MBP-fused magainin in E. coli decreased with the increasing size of multimers possibly because of the transcription and translation inhibition by the multimeric peptides. After purification using an amylose affinity column, the fusion protein was digested by factor Xa at a specific cleavage site between the monomers. The recombinant magainin had an antimicrobial activity identical to that of synthetic magainin. This experiment shows that a biologically active, antimicrobial peptide magainin can be produced by fusing to MBP, along with a promoter inversion vector system.

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Enoxaparin as an Anticoagulant in a Multipara with a Mechanical Mitral Valve: A Case Report

  • Yo Seb Lee;Jun Seok Kim
    • Journal of Chest Surgery
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    • 제56권6호
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    • pp.452-455
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    • 2023
  • Patients who have undergone mechanical valve replacement require anticoagulation therapy with warfarin to prevent thromboembolism. However, administering warfarin to pregnant patients increases their risk of warfarin embryopathy or central nervous system disorders. Consequently, safer alternatives, such as heparin or low-molecular-weight heparin injection, are substituted for warfarin. However, limited research has been conducted on this subject, with no large-scale studies and particularly few investigations involving multiparous patients. A patient who had previously undergone mechanical mitral valve replacement for atrial septal defect and mitral stenosis received anticoagulant therapy with enoxaparin during 2 pregnancies. Upon confirmation of pregnancy, warfarin was replaced with subcutaneously injected enoxaparin with a dosage of 1 mg/kg at 12-hour intervals. The enoxaparin dosage was controlled using an anti-factor Xa assay, with a target range of 0.3-0.7 IU/mL. Intravenous heparin injections were administered starting 3 days prior to the expected delivery date and were continued until delivery, after which warfarin was resumed. No complications were observed during the deliveries.

치환리터럴에 의한 Quaternary Galois Field Sum-Of-Product(QGFSOP)형 1-변수 함수의 합성과 실현 (Syntheses and realization of Quaternary Galois Field Sum-Of-Product(QGFSOP) expressed 1-variable functions Permutational Literals)

  • 박동영;김백기;성현경
    • 한국항행학회논문지
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    • 제14권5호
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    • pp.710-717
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    • 2010
  • Quaternary 논리에서 생성 가능한 1-qudit(1-variable quantum digit) 함수는 총 256개가 존재하지만 이들 중에서 가장 유용한 것은 "0,1,2,3"의 치환에 의해 $Ax^C$+D(GF4)형의 QGFSOP 표현이 가능한 24개이다. 본 논문에서는 24개 1-qudit 함수들의 $Ax^C$+D(GF4) 연산에서 피연산자인 피승수 A와 피기수 D를 다단 종속된 치환리터럴의 제어인자로 사용하는 치환리터럴(Permutational Literals, PL) 표현과 QPL(Quaternary PL) gate를 제안하였다. 그리고 상호치환 'ab', 가산 '+D', 그리고 승산 'xA'과 같은 세 개의 PL 연산자를 사용하여 QGFSOP 표현된 24개 (1-qudit) 함수를 합성하기 위한 PL 합성법을 제안하였다. 끝으로 PL 합성법을 실현하기 위한 $Ax^C$+D(GF4) 구조와 연산회로 및 CMOS 실현 방법을 제시하였다.

암환자의 정맥혈전색전증 치료에서 rivaroxaban과 dalteparin의 출혈 부작용 비교 (Comparisons of Bleeding Risks between Rivaroxaban and Dalteparin for Treatment of Venous Thromboembolism in Cancer Patients)

  • 김윤경;안숙희;김재연;정지은;곽혜선
    • 한국임상약학회지
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    • 제26권3호
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    • pp.195-200
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    • 2016
  • Background: Venous thromboembolism (VTE) is a common and life-threating condition in cancer patients. Low molecular weight heparins (LMWH), such as dalteparin, are recommended in the treatment of VTE. Also, rivaroxaban, an orally administered direct factor Xa inhibitor, was approved for the treatment of VTE. It showed similar efficacy to standard therapy (LMWH or warfarin) and was associated with significantly lower rates of major bleedings. However, in the real world, bleeding has been reported to occur frequently in cancer patient receiving rivaroxaban. The goal of this research was to analyze bleeding risks between rivaroxaban and dalteparin for treatment of VTE in cancer patients. Methods: Medical records of oncology patients who were treated with rivaroxaban or dalteparin for VTE from July 2012 to June 2014 were retrospectively reviewed. Data collected were as follows: age, sex, weight, height, cancer types and stages, ECOG (eastern cooperative oncology group) PS (performance score), VTE types, concurrently used medications, study drug information (dose and duration of therapy), INR (international normalized ratio), PT (prothrombin time), and platelet counts. Bleeding was classified into major bleedings, clinically relevant non-major bleedings, and minor bleedings. Results: A total of 399 patients were included in the study. Of these patients, 246 were treated with rivaroxaban and 153 with dalteparin. Bleeding rates were significantly higher in the rivaroxaban group than in the dalteparin group (adjusted odds ratio (AOR) 2.09, 95% CI 1.22-3.60) after adjusting for confounders. In addition, rivaroxaban remained independently associated with 1.78-fold (95% CI 1.14-2.76) shorter time to bleeding compared to dalteparin after adjusting other factors known to be associated with poor outcomes. Conclusion: This study suggested that rivaroxaban was associated with an increased risk of bleedings in cancer patients.