• 제목/요약/키워드: proteinase

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Discovery of New Proteinase Inhibitor for the Treatment of Osteoporosis

  • 손문호
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 2001년도 춘계학술대회
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    • pp.89-99
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    • 2001
  • ■ Cathepsin K is a attractive target for selectively and efficiently modulating the osteoclastic bone resorption. ■ OST-1857 is a lead compound which is specifically targeted to cathepsin K and showed efficacy in TPTX rats. ■ OST-compounds are in process of the preclinical study, joined by Yuhan research center.

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An EST survey of genes expressed in liver of rock bream(Oplegnathus fasciatus) with particular interests on the stress-responsive and immune-related genes

  • Park, Byul-Nim;Park, Ji-Eun;Kim, Ki-Hong;Kim, Dong-Soo;Nam, Yoon-Kwon
    • 한국양식학회:학술대회논문집
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    • 한국양식학회 2003년도 추계학술발표대회 논문요약집
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    • pp.43-43
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    • 2003
  • EST analysis was performed to identify stress-responsive and immune-related genes from rock bream (Oplegnathus fasciatus). cDNA libraries were constructed with liver and randomly chosen 624 clones were subjected to automated sequence analysis. Of 624 clones sequenced in total, approximately 15% of ESTs was novel sequences (no match to GenBank) or sequences with high homology to hypothetical/unknown genes. The bioinforamtic sequence analysis including functional clustering, homology grouping, contig assembly with electronic northern and organism matches were carried out. Several potential stress-responsive biomarker and/or immune-related genes were identified in all the tissues examined. It included lectins, ferritins, CP450, proteinase, proteinase inhibitors, anti-oxidant enzymes, various heat-shock proteins, warm temperature acclimation protein, complements, methyltransferase, zinc finger proteins, lysozymes, macrophage maturation associated protein, and others. This information will offer new possibilities as fundamental baseline data for understanding and addressing their molecular mechanism involved in host defense and immune systems of this species.

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Endochondral Ossification Signals in Cartilage Degradation During Osteoarthritis Progression in Experimental Mouse Models

  • Kawaguchi, Hiroshi
    • Molecules and Cells
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    • 제25권1호
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    • pp.1-6
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    • 2008
  • Osteoarthritis (OA), one of the most common skeletal disorders characterized by cartilage degradation and osteophyte formation in joints, is induced by accumulated mechanical stress; however, little is known about the underlying molecular mechanism. Several experimental OA models in mice by producing instability in the knee joints have been developed to apply approaches from mouse genetics. Although proteinases like matrix metalloproteinases and aggrecanases have now been proven to be the principal initiators of OA progression, clinical trials of proteinase inhibitors have not been successful for the treatment, turning the interest of researchers to the upstream signals of proteinase induction. These signals include undegraded and fragmented matrix proteins like type II collagen or fibronection that affects chondrocytes through distinct receptors. Another signal is proinflammatory factors that are produced by chondrocytes and synovial cells; however, recent studies that used mouse OA models in knockout mice did not support that these factors have a role in the central contribution to OA development. Our mouse genetic approaches found that the induction of a transcriptional activator Runx2 in chondrocytes under mechanical stress contributes to the pathogenesis of OA through chondrocyte hypertrophy. In addition, chondrocyte apoptosis has recently been identified as being involved in OA progression. We hereby propose that these endochondral ossification signals may be important for the OA progression, suggesting that the related molecules can clinically be therapeutic targets of this disease.

Comparative Quantification of LacZ (β-galactosidase) Gene from a Pure Cultured Escherichia coli K-12

  • Han, Ji-Sun;Kim, Chang-Gyun
    • Environmental Engineering Research
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    • 제14권1호
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    • pp.63-67
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    • 2009
  • Escherichia coli K-12 (E. coli K-12) is a representative indicator globally used for distinguishing and monitoring dynamic fates of pathogenic microorganisms in the environment. This study investigated how to most critically quantify lacZ ($\beta$-galactosidase) gene in E. coli K-12 by two different real-time polymerase chain reaction (real-time PCR) in association with three different DNA extraction practices. Three DNA extractions, i.e., sodium dodecyl sulfate (SDS)/proteinase K, magnetic beads and guanidium thiocyanate (GTC)/silica matrix were each compared for extracting total genomic DNA from E. coli K-12. Among them, GTC/silica matrix and magnetic beads beating similarly worked out to have the highest (22-23 ng/${\mu}L$) concentration of DNA extracted, but employing SDS/proteinase K had the lowest (10 ng/${\mu}L$) concentration of DNA retrieved. There were no significant differences in the quantification of the copy numbers of lacZ gene between SYBR Green I qPCR and QProbe-qPCR. However, SYBR Green I qPCR obtained somewhat higher copy number as $1{\times}10^8$ copies. It was decided that GTC/silica matrix extraction or magnetic beads beating in combination with SYBR Green I qPCR can be preferably applied for more effectively quantifying specific gene from a pure culture of microorganism.

모기 살충성 Bacillus thuringiensis 21-2균주의 용혈성 내독소 단백질의 특성 (Characteristics of Hemolysin in Mosquitocidal Bacillus thuringiensis strain 21-2)

  • 김광현;김위종;김영희;김병우
    • 한국미생물·생명공학회지
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    • 제30권3호
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    • pp.230-234
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    • 2002
  • 모기 살충성 Bacillus thuringiensis subspangiensis 21-2균주의 용혈성 내독소 단백질의 특성을 검토하고자 21-2균주의 용혈성을 가진 유전자를 Escherichia coli HBIO쎄 형질전환시켰다. 이들 중에서 형질전환 균주 47은 독소 단백질을 생산하며 2.5 kb DNA을 함유한다는 것을 효소항체법, immunoblot및 DNA전기영동법으로 확인하였다. 또한, 형질전환 균주 47-5는 2.5 kb DNA를 다시 Hind ll견 절단하여 pUC118 연결시켜서 조제하였다 그 결과형질전환 균주 47-5은 1.Bkb DNA를 함유하며, 23 kD꺼 독소 단백질을 발현하고, 발현된 독소 단백질은 Aedes aegypti모기 유충에 독성을 나타내었다. 또한 23 kDa의 내독소 단백질 그 자체로는 사람의 적혈구를 용해하지 못하였으나, proteinase K로 처리한 후에는 적혈구에 대해 용혈성을 나타내었다.

Inhibition of Various Proteases by MAPI and Inactivation fo MAPI by Trypsin

  • Lee, Hyun-Sook;Kho, Yung-Hee;Lee, Kye-Joon
    • Journal of Microbiology and Biotechnology
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    • 제10권2호
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    • pp.181-186
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    • 2000
  • MAPI (microbial alkaline protease inhibitor) was isolated from cultrue broth of Streptomyces chromofuscus SMF28. The Ki values of MAPI for the representative serine proteases such as chymotrypsin and proteinase K were 0.28 and $0.63{\;}\mu\textrm{M}$, respectively, and for the cysteine proteases cathepsin B and papain were 0.66 and $0.28{\;}\mu\textrm{M}$, respectively. These data indicate that MAPI is not a potent selective inhibitor of serine or cysteine proteases. Progress curves for the inhibition of three proteases by MAPI exhibithe characteristic patterns; MAPI exhibited slow-binding inhibition of cathepsin B. It was rapidly associated with chymotrypsin before the addition of substrate and then reactivation of MAPI-inhibited enzyme was investigated in the presence of substrate. On the other hand, MAPI-proteinase K interaction was typical for those classical inhibitors. When MAPI was incubated with trypsin, there was an extensive reduction in the ingibitory activities of MAPI corresponding to 66.5% inactivation of MAPI, indicating that trypsin-like protease may play a role in the decrease of the inhibitory activity during cultivation.

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Single Somatic Embryogenesis from Transformant with Proteinase II Gene in Panax ginseng C.A. Meyer

  • Yang, Deok-Chun;Kim, Se-Young;Rho, Yeong-Deok;Kim, Moo-Sung
    • Plant Resources
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    • 제6권3호
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    • pp.205-210
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    • 2003
  • Ginseng(Panax ginseng C.A. Meyer) is a perennial herbaceous plant which grows very slowly. It takes about 3 to 4 years from seeding to collecting the ripe seeds and the ginseng propagation is very difficult. and so, it is very difficult to breed ginseng plant. Ginseng tissue culture was started from at 1960, and ginseng commercial product by in vitro callus culture was saled, however upto now, regenerants were not planted to soil normally. Recently, plant genetic engineering to produce transgenic plants by introducing useful genes has been advanced greatly. In a present paper, transformation of ginseng plants was achieved by co-cultivation with Agrobacterium harboring the binary vector coding Proteinase-II gene, which confer resistant or tolerant to insect pests, The binary vector for transformation was constructed with disarmed Ti-plasmid and with double 35S promoter. The NPT II gene and introduced genes of the transgenic ginseng plants were successfully identified by the PCR. Especially the transgenic ginseng plants were regenerated using new techniques such as repetitive single somatic embryogenesis.

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Cytokinins overproduction에 따른 담배형질전환체의 변화 (Phenotypic Alterations in Transgenic Tobacco Plants that Overproduce Cytokinins)

  • 정용윤
    • 자연과학논문집
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    • 제10권1호
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    • pp.33-37
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    • 1998
  • 식물의 주요 phytohormone의 하나인 cytokinin은 식물체의 줄기와 뿌리성장 그 외에도 영양의 전달이나 노화방지, 열매숙성 등 식물의 성장과 발달에 미치는 영향은 크고 다양하다. Cytokinin 생합성에 관여하는 효소를 생산하는 것으로 알려져 있으며 토양박테리아 Agrobacterium tumefaciens에 존재하는 유전자인 isopentenyl transferase (jpt)를 이용한 많은 분자생물학적 연구가 진행되어 왔는데 그 중 하나로 이 연구에서 jpt 유전자에 의한 cytokinin의 overproduction이 식물체에 성장과 발달에 어떠한 영향을 주는지 관찰하고 그 결과가 제시할 수 있는 작물의 유전 공학적 이용가능성에 대하여 알아본다.

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Bacillus subtilis 168 균주가 분비하는 5 kDa 크기의 Bacteriocin (A Bacteriocin of 5-kDa in Size Secreted by Bacillus subtilis 168)

  • 권건희;이황아;김정환
    • 한국미생물·생명공학회지
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    • 제38권2호
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    • pp.163-167
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    • 2010
  • B. subtilis 168 균주는 배양중 항균물질을 배지중으로 분비하며 배양상등액은 몇몇 그램 양성균을 저해한다. B. cereus와 L. monocytogenes의 저해 정도가 가장 컸었다. 배양상등액을 proteas와 proteinase K로 처리할 경우 항균력이 상실되어서 항균물질은 단백질성(박테리오신) 임을 알수있었다. Tricine SDS-PAGE에 의해서 박테리오신 분자량은 5 kDa으로 확인되었다. 박테리오신은 민감한 균을 죽임으로써 생육을 저해하는 것으로 밝혀졌다. 이상 결과들에서 B. subtilis 168은 청국장과 같은 B. cereus 오염이 문제되는 발효식품들의 종균으로 유용할 것으로 생각된다.