• 제목/요약/키워드: Factor 9

검색결과 9,410건 처리시간 0.037초

인체 혈액응고 9인자 cDNA cloning 및 Escherichia coli 에서의 발현 (Cloning and Expression of Human Clotting Factor 9 cDNA un Escherichia coli)

  • Young Won Lee;Hyang Suk Hur;Myoung Hee Kim
    • 대한의생명과학회지
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    • 제2권2호
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    • pp.231-240
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    • 1996
  • 인체 혈액 응고 9인자는 간에서 생성되며 461개의 아미노산으로 구성된 당 단백질이다. 따라서 인체 혈액 응고 9인자 cDNA를 찾기 위해 태아의 간(fetal liver) cDNA library를 PCR(Polymerase Chain reaction) 방법으로 screening하였으며, 그 결과 ATG개시 코돈으로부터 TAA종료 코돈까지 포함하는 1.4 kb의 9인자 cDNA를 찾았다. 또한 클론된 9인자 cDNA를 박테리아에서 발현시키기 위해 박테리아 발현 벡터인 pGEX-2T 플라스미드에 클로닝하므로써 pGEX-F9 플라스미드를 제조하였다. pGEX-F9로 형질전환된 E. coli에서 PGEX-F9의 발현을 유도하면 73 kDa 크기의 GST-factor9 융합 단백질이 다량생성되며 , 이 단백질이 혈액 응고 9인자 단백질을 함유하는 융합 단잭질임을 혈액 응고 9인자 항체를 이용한 Western blot으로 입증하였다. E. coli에서 발현된 GST-factor 9 융합 단백질은 전체 단백질의 약 20%를 차지하며 GST agarose bead를 이용한 one step purificarion 방법을 통해 GST-factor9 융합 단백질을 쉽게 분리 할 수 있다.

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Development of a Highly Active Fluorescence-Based Detector for Yeast G Protein-Coupled Receptor Ste2p

  • Hong, Jin Woo;Ahn, Hee Jun;Baek, Jee Su;Hong, Eun young;Jin, Dong Hoon;Khang, Yong Ho;Hong, Nam Joo
    • Journal of Microbiology and Biotechnology
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    • 제28권10호
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    • pp.1589-1603
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    • 2018
  • Twenty analogs of $[Orn^6,D-Ala^9]{\alpha}-factor$ were synthesized and assayed for their biological activities: seven analogs of $[Orn^6,X^9]{\alpha}-factor$, seven analogs of $[X^6,D-Ala^9]{\alpha}-factor$, five analogs of $[X^5,X^6,D-Ala^9]{\alpha}-factor$, and native ${\alpha}-factor$ (X = amino acids). Their biological activities (halo, gene induction, and affinity) were measured using S. cerevisiae Y7925 and LM102 and compared with those of native ${\alpha}-factor$ (100%). G protein-coupled receptor was expressed in strain LM102 containing pESC-LEU-STE2 vector. $[Dap^6,D-Ala^9]{\alpha}-factor$ with weak halo activity (10%) showed the highest receptor affinity (> 230%) and the highest gene induction activity (167%). $[Arg^6,D-Ala^9]{\alpha}-factor$ showed the highest halo activity (2,000%). The number of active binding sites per cell (about 20,000 for strain LM102) was determined using a newly-designed fluorescence-based detector, $[Arg^6,D-Ala^9]{\alpha}-factor-Edan$, with high sensitivity (12,500-fold higher than the absorption-based detector $[Orn^6]{\alpha}-factor-[Cys]_3$).

공동주택의 분양가 결정을 위한 영향요인 도출 및 중요도 분석 (The Analysis of Importance Ratio Factors of Sales Price for Apartment Building Construction Projects)

  • 양옥희;김민석;황욱선;김용수
    • 한국건설관리학회논문집
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    • 제12권5호
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    • pp.127-136
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    • 2011
  • 본 연구는 공동주택의 분양가 결정을 위한 영향요인 도출 및 중요도 분석을 목적으로 수행되었다. 이를 위해 서울, 경기, 충청지역별 각각 3곳의 공동주택을 사례대상으로 선정하였으며, 관련 연구자료 및 전문가 면담을 통해 분양가 영향요인을 도출하였다. 이렇게 도출된 분양가 영향요인을 대상으로 각 지역별 전문가 설문을 통해 AHP기법을 활용하여 분양가 영향요인의 중요도를 분석하였다. 상기와 같은 목적과 방법에 따라 진행된 본 연구의 결론을 요약하면 다음과 같다. 1) 공동주택 분양가 영향요인은 원가, 단지, 주택, 교통, 지역, 토지, 정책 투자가치요인의 총 8가지 항목으로 도출되었다. 2) 각 지역별 영향요인의 중요도 분석결과를 정리하면 다음과 같다. 서울지역은 투자가치요인(16.0%), 정책요인(14.6%), 원가요인(13.9%)이 주요인으로 선정되었으며, 경기지역은 지역요인(15.9%) 교통요인(13.8%) 투자가치요인(13.4%)이 주요인으로 선정되었다. 충청지역은 지역요인(17.1%), 교통요인(16.4%), 주택요인(14.9%)이 주요인으로 선정되었다.

Highly Active Analogs of α-Factor and Their Activities Against Saccharomyces cerevisiae

  • Ahn, Hee Jun;Hong, Eun Young;Jin, Dong Hoon;Hong, Nam Joo
    • Bulletin of the Korean Chemical Society
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    • 제35권5호
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    • pp.1365-1374
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    • 2014
  • Thirteen analogs of tridecapeptide ${\alpha}$-factor (WHWLQLKPGQPMY) of Saccharomyces cerevisiae with C- or N-terminal Trp extension and isosteric replacement by Aib at position 8 and 11, Trp at position 13, D-Ala at position 9, and Orn and Glu at position 6 were synthesized and assayed for their biological activity. Receptor binding assay was carried out using our newly developed spectrophotometric method with detector peptide 14. C- or N-terminal extended analogs, ${\alpha}$-factor-$[Trp]_n$ (n =1-5) 1-5 and $[N-Trp]_1$-${\alpha}$-factor 6, were all less active than native ${\alpha}$-factor and gradual decreases in both activity and receptor affinity were observed with greater Trp extension. Trp-substituted analog at position 13, $[Trp^{13}]{\alpha}$-factor 7, exhibited about 2-fold reductions in both activity and receptor affinity. Aib-substituted analogs, $[Aib^8]{\alpha}$-factor 8 and $[Aib^{11}]{\alpha}$-factor 9, showed 5- to 10-fold reduction in activity as well as 3-fold reduction in receptor affinity compared to native ${\alpha}$-factor. $[Orn^6]{\alpha}$-factor 10 demonstrated strong potency with a 7.0-fold increase in halo activity as well as 1.8-fold increase in receptor affinity compared to native ${\alpha}$-factor. For two double substituted analogs, [$Glu^6,{\small{D}}-Ala^9$]${\alpha}$-factor 12 showed the slightly decreased potency in halo activity compared to analog 10, whereas [$Orn^6,{\small{D}}-Ala^9$]${\alpha}$-factor 11 exhibited 15-fold higher halo activity as well as nearly 3-fold higher receptor affinity compared to native ${\alpha}$-factor.

RAW 264.7 세포에서 sulforaphane의 파골세포형성 저해효과 (Effects of Sulraphane on Osteoclastogenesis in RAW 264.7)

  • 황준호;이미란;강창희;부희정
    • 농업생명과학연구
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    • 제50권2호
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    • pp.151-160
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    • 2016
  • 염증성 사이토카인은 파골세포형성과정에서 중요한 요인이며, 뼈의 흡수는 자주 골다공증과 연결된다. 설포라판은 보로콜리의 화뢰로 부터 분리된 물질로 염증성 사이토카인을 억제한다고 알려져 있다. 본 실험에서는 Receptor activator of nuclear factor kappaB ligand(RANKL)로 자극된 세포에서 설포라판이 파골세포 형성 억제에 대한 효과를 측정하였다. 설포라판은 대식세포인 RAW 264.7 세포에서 파골세포 특이 마커 유전자인 tartrate-resistant acid phosphatase(TRAP), Cathepsin K, matrix metalloproteinase 9(MMP-9), calcitonin receptor을 저해하였으며, TRAP, MMP-9, tumor necrosis factor receptor-associated factor 6(TRAF6)와 전사인자인 nuclease factor of activated T cells(NFATc1)의 단백질 발현과 RANKL로 자극하였을 때 전자인사인 nuclear factor kappaB(NF-kappaB)의 전사활성도 억제 하였다. 이와 같은 결과로 설포라판이 NF-kappaB의 전사활성 억제뿐만 아니라, 파골세포형성인자(TRAP, cathepsin K, MMP-9, calcitonin, NFATc1)와 NFATc1의 발현을 억제시키는 효과가 있음을 확인하였다.

생쥐 난소의 발생단계에 따른 Growth Differentiation Factor-9의 유전자 발현 (Expression of Growth Differentiation Factor-9 in the Mouse Ovaries at Different Developmental Stages)

  • 윤세진;이경아;고정재;차광열
    • 한국발생생물학회지:발생과생식
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    • 제3권1호
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    • pp.95-100
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    • 1999
  • Growth/differentiation factor-9 (GDF-9)은 transforming growth factor $\beta$ (TGF-$\beta$) superfamily의 member로서 난소의 난자에서만 특이적으로 발현되며 정상적인 난포발달에 있어 필수적인 성숙인자로 최근에 알려졌다. 본 연구는 RT-PCR을 통해 생쥐의 원시난포에서의 GDF-9 mRNA의 발현 여부와 함께 난포의 발달단계에 따른 상대적인 발현량을 분석하고자 실시하였다. 본 실험에는 ICR 생쥐를 사용하여 질전 (vaginal plug)이 확인된 날을 1일로 하여 임신 19일의 태아와 태어난 날을 1일로 하여 생후 1일, 10일, 21일, 28일된 생쥐 난소를 실험에 사용하였다. 각 발달단계의 난소조직으로부터 total RNA를 추출하여 GDF-9 유전자 발현 여부를 확인하였으며 이들을 $\beta$-actin에 대해 상대적인 정량분석을 하였다. GDF-9 유전자 발현은 아직은 성장을 시작하지 않은 임신 19일의 태아의 난소, 대부분이 원시난포로 이루어진 태어난 날의 생쥐 난소에서도 확인되었으며, 성장이 왕성하게 진행되고 있는 난포 즉, antrum 형성 이전의 growing follicles이 주를 이루는 생후 10일째의 난소에서 가장 높은 GDF-9 유전자 발현이 관찰되었다. 나머지 단계의 난소에서는 거의 비슷한 정도로 발현함을 관찰할 수 있었다. 본 연구의 결과는 생쥐의 원시난포에도 GDF-9 transcript가 존재한다는 것을 확실하게 증명하였으며, GDF-9이 생쥐의 초기 난포발달에 중요한 역할을 할 것이라는 가능성을 시사한다.

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Mangiferin inhibits tumor necrosis factor-α-induced matrix metalloproteinase-9 expression and cellular invasion by suppressing nuclear factor-κB activity

  • Dilshara, Matharage Gayani;Kang, Chang-Hee;Choi, Yung Hyun;Kim, Gi-Young
    • BMB Reports
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    • 제48권10호
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    • pp.559-564
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    • 2015
  • We investigated the effects of mangiferin on the expression and activity of metalloproteinase (MMP)-9 and the invasion of tumor necrosis factor (TNF)-$\alpha$-stimulated human LNCaP prostate carcinoma cells. Reverse-transcription polymerase chain reaction (RT-PCR) and western blot analysis showed that mangiferin significantly reversed TNF-$\alpha$-induced mRNA and protein expression of MMP-9 expression. Zymography data confirmed that stimulation of cells with TNF-$\alpha$ significantly increased MMP-9 activity. However, mangiferin substantially reduced the TNF-$\alpha$-induced activity of MMP-9. Additionally, a matrigel invasion assay showed that mangiferin significantly reduced TNF-$\alpha$-induced invasion of LNCaP cells. Compared to untreated controls, TNF-$\alpha$-stimulated LNCaP cells showed a significant increase in nuclear factor-${\kappa}B$ (NF-${\kappa}B$) luciferase activity. However, mangiferin treatment markedly decreased TNF-$\alpha$-induced NF-${\kappa}B$ luciferase activity. Furthermore, mangiferin suppressed nuclear translocation of the NF-${\kappa}B$ subunits p65 and p50. Collectively, our results indicate that mangiferin is a potential anti-invasive agent that acts by suppressing NF-${\kappa}B$-mediated MMP-9 expression.

불임증(不姙症) 환자(患者)의 통계적(統計的) 고찰(考察);서울대학교병원(大學校病院) 불임상담실(不姙相談室) 1872 예(例)의 분석(分析) (An Analysis of Infertility Patients)

  • 장윤석;이진용;문신용;김정구;최승헌;임용택
    • Clinical and Experimental Reproductive Medicine
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    • 제12권1호
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    • pp.47-70
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    • 1985
  • This study was presented of the 1,872 cases of infertile couples who visited and examined at the sterility clinic of Department of Obstetrics & Gynecology, Seoul National University Hospital from Sept., 1980 to Dec., 1983. Age, duration of infertility, past medical history, and other general factors were analyzed, and the factors responsible for infertility were classified and discussed. Mode of treatment, outcome of pregnancy, pregnancy rate responsible for each factor were also presented. The results were as follows: 1) The infertility was primary in 1,128, or 60.3% and secondary in 744, or 39.7%. 2) The age between 26 and 30 years of age comprised about one half of the total patients. 3) The duration of infertility between 1 and 4 years comprised about three quarters of the total patients, and the mean duration was 3.8 years. 4) The most common medical history in primary infertility was tuberculous disease, and that in secondary infertility was history of previous laparotomy. 5) About two thirds of antecedent pregnancies were abortion. 6) The major etiologic factor of infertility were male factor in 12.3%, tubal factor in 38.8%, ovulatory failure in 25.4%, uterine factor in 8.8%, cervical factor in 5.2%, peritoneal factor in 9.5%, and no demonstrable cause in 11.3%. 7) The types of male factor were azoospermia in 61.6%, oligospermia in 25.8%, low motility in 11.6%, and other abnormality in 1.0%. 8) The types of ovulatory failure were ovarian failure in 7.4%, hypothalamo-pituitary failure in 8.1 %, hypothalamo-pituitary dysfunction (including Polycystic ovarian syndrome) in 30.2%, and hyperprolactinemia in 22.4%. 9) The types of uterine factor were endometrial tuberculosis in 27.5%, uterine synechia in 33.8%, uterine anomaly in 19.7%, myoma and polyp in 9.1 %, and luteal phase defect in 9.9%. 10) The types of peritoneal factor were pelvic adhesion in 80.9% and endometriosis in 19.6%. 11) Surgeries were done in 408 patients, and they were salpingolysis, lysis of extraadnexal adhesion, salpingostomy, fimbrioplasty, ovarian wedge resection for polycystic ovarian disease, tubo-tubal anastomosis, and tubo-uterine implantation in orders. 12) 243 pregnancies were achieved during the infertility work-up, of which livebirth was 46.5%, ectopic pregnancy was 7.4%, spontaneous abortion was 7.8%, and on-going pregnancy or lost to follow-up was 36.2%. 13) Pregnancy rates in various factors were male factor in 18.7%, ovulatory factor in 31.7%, tubal factor in 24.2%, uterine factor in 34.6%, cervical factor in 19.0%, peritoneal factor in 29.0%, combined factors in 10.5%, and unexplained infertility in 37.1%. Pregnancy rate in whole patients was 25.2%.

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