• Title/Summary/Keyword: E1B-19k

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Molecular Cloning and Analysis of Nucleotide Sequence of Xylanase Gene (xynk) from Bacillus pumilus TX703 (Bacillus pumilus TX703 유래 Xylanase 유전자(xynK)의 Cloning과 염기서열 분석)

  • 박영서
    • Journal of Life Science
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    • v.12 no.2
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    • pp.188-199
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    • 2002
  • A gene coding for xylanase from thermo-tolerant Bacillus pumilus TX703 was cloned into Escherichia coli DH5 $\alpha$ using pUC19. Among 7,400 transformants, four transformants showed clear zones on the detection agar plates containing oat-spells xylan. One of them which showed highest xylanase activity was selected and its recombinant plasmid, named pXES106, was found to carry 2.24 kb insert DNA fragment. When the nucleotide sequence of the cloned xylanase gene (xynK) was determined, xynK gene was found to consist of 1,227 base-pair open reading frame coding for a polypeptide of 409 amino acids with a deduced molecular weight of 48 kDa. The coding sequence was preceded by a putative ribosome binding site, the transcription initiation signals, and cia-acting catabolite responsive element. The deduced amino acids sequence of xylanase is similar to those of the xylanases from Hordeum vulgare (barley) and Clostridium thermocellum, with 39 and 31% identical residues, respectively. The amino acids sequence of this xylanase was quite different from those of the xylanases from other Bacillus species.

The Crystal and Molecular Structure of Dipropargyldiphenylmethane (디프로파질디페닐메탄의 결정 및 분자구조)

  • Ahn Choong Tai;Choi Sam-Kwon
    • Journal of the Korean Chemical Society
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    • v.37 no.5
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    • pp.473-476
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    • 1993
  • Dipropargyldiphenylmetane, $C_{19}H_{16}, crystallizes in a monoclinic space group $C2/_c$$ with a = 11304(3), b = 20.799(5), c = 6.622(2)${\AA}$, ${\beta} = 112.8(3)^{\circ}$, Z = 4, V = 1435.3${\AA}^3,\;F(000)\;=\;520,\;D_c\;=\;1.14g{\cdot}cm^{-3}$ and ${\mu}\;=\;0.32\;cm^{-1}$. The structure was solved by direct methods and all non-H atoms were identified in the E-map. The final refinement gave R = 0.055 from 1328 unique observed reflections with I $\geq$ -1.0 $\sigma(I).$ The molecule belongs to the point group $C_2$ of Symmetry by possessing the 2-fold axis which coincides witeh the crystallographic symmetry axis in the unit cell. The linear propargyl moiety is nearly $perpendicular(94.2)^{\circ}$ to the molecular plane of the benzene ring. The internal angle of methane carbon atoms in $108.1(1)^{\circ}$, bonding to the benzene and the propargyl moiety with the bond lengths of 1.530(2) and $1.560(2)\AA$, respectively. The shortest contant between the molecules is $3.538(2)\AA$ between C(9) and C(9) (-x, y, -1/2-z).

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Coloration of Synthetic fiber fabrics with Loess(I) (합성섬유 직물에 대한 황토염색(I))

  • Lee, Jeon-Sook;Choi, Kyung-Eun
    • Korean Journal of Human Ecology
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    • v.8 no.1
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    • pp.19-24
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    • 2005
  • This paper is to know the possibility of coloration of synthetic fibers with loess. Fabrics woven with polyester, nylon, and acrylic fibers were dyed by a dip-pad operation with a laboratory mangle. Drying, curing at 180$^{\circ}C$ for 10 minutes and washing were followed. Effect of curing after drying were investigated in terms of K/S values and SEM microphotographes. Reddish-yellow(a:75.13, B:15.14, ${\Delta}$E:45.31) colored fabric ascertain possibility of coloration of synthetic fibers with loess. Acrylic fiber shows highest dye uptake of the three kinds of fibers and fastness increases by curing after drying.

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Preparation and Evaluation of Mutivitamin Emulsion (복합비타민 유제의 제조와 평가)

  • Lee, Moon-Seok;Cho, Hea-Young;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.32 no.1
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    • pp.13-19
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    • 2002
  • Water-lipid soluble multivitamin formulations were widely used to reduce the disease and stress of animals as husbandry has made a remarkable progress in recent. But the efficiency of these formulations is far from satisfactory. So, this study was attempted to develop the physically and chemically stable and useful multivitamin o/w emulsion. Multivitamin o/w emulsion composed of water, soybean oil (10%, v/v), vitamin A, D, E, K, $B_2,\;B_6,\;B_{12}$ and panthenol. To make a stable o/w emulsion, the egg lecithin (2%, w/v) and glycerin (2.5%, w/v) were used for emulsifier and thickening agent, respectively. The oil in water emulsion system was manufactured by microfluidizer and the physicochemical stability of this emulsion was evaluated. The average particle size and interfacial tension were measured. From the result of interfacial tension tested, critical micelle concentration of the egg lecithin was 0.5% (w/v) and optimal concentration for the preparation of emulsion was 2% (w/v). The mean particle size was about $0.6\;{\mu}m$ which was suitable for injections. Short-term accelerated stability as physical stability study was tested by centrifuging and freeze-thawing the emulsion samples. The additions of vitamins resulted in the increment of particle size and reduction of physical stability of emulsion. But it is not an enormous problem for the stability of emulsion. Also, we have performed the long-period preservation stability test for the vitamins. All vitamins were analysed by HPLC. The result of storage under $4^{\circ}C$ and dark conditions demonstrated that all vitamins were maintained stable at least 16 weeks, except for vitamin $B_{12}$.

A Study on the Effect of Herbal-acupulcture with Angelicae Pubescentis Radix Solution at Joksamni$(ST_{36})$ on Collagen-induced Arthritis (족삼리(足三里) 독활약침(獨活藥鍼)이 Collagen-induced Arthritis에 미치는 영향)

  • Yang, Gi-Young;Kim, Young-Il;Lee, Hyeon
    • Journal of Acupuncture Research
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    • v.23 no.3
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    • pp.191-206
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    • 2006
  • Objective & Methods : The purpose of this study is to observe the effects of Angelicae Pubescentis Radix herbal-acupuncture solution(APR-HAS) at Joksamni(ST36) on collagen IT induced arthritis in DBA/1J mice. The author performed several experimental items to analyze several cytokines and immune cells related with RA. Results : 1. In the APR-HA group, the incidence of arthritis and arthritis index were significantly decreased. 2. In APR-HA group, the levels of IL-6, $INF-{\gamma}$, $TNF-{\alpha}$, IgG, IgM, $IL-{\beta}$ and Anti-collagen II in serum of the CIA mouse were significantly decreased. 3. In APR-HA group, the level of $IFN-{\gamma}$, IL-4 in the CIA mouse spleen cell culture were significantly decreased. 4. In histology, the cartilage destruction and synovial cell proliferation were decreased in the APR-HA group, and the collagen fiber expressions in the APR-HA group were similar with that of the Normal group. 5. In the APR-HA group, CD3e+/CD19+ and CD4+/CD8+ were similarly maintained as Normal group in the CIA mouse lymph nodes, 6. In the APR-HA group, CD3e+/CD69+ was significantly decreased in the CIA mouse joint. 7. In the APR-HA group, CD11a+/CD19+ and CD11b+/Gr-l+ were significantly decreased in the CIA mouse lymph nodes 8. In the APR-HA group, CD4+/CD25+ was decreased in the CIA mouse spleen cell. 9. In the APR-HA group, CD4+/CD25+ was similarly maintained as Normal group in the CIA mouse lymph nodes.

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Development of a Virus Elution and Concentration Procedure for Detecting Norovirus in Oysters

  • Ha, Sook-Hee;Woo, Gun-Jo;Hwang, In-Gyun;Choi, Weon-Sang
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1150-1154
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    • 2009
  • Low levels of virus contamination and naturally occurring reverse transcription-polymerase chain reaction (RT-PCR) inhibitors restrain virus detection in oysters. A rapid and efficient oyster-processing procedure that can be used for sensitive virus detection in oysters was developed. Poliovirus type 1 Sabin strain was used to evaluate the efficacy of virus recovery. The procedure included (a) acid-adsorption and elution with buffers (0.25M glycine-0.14 M NaCl, pH 7.5; 0.25M threonine-0.14M NaCl, pH 7.5); (b) polyethylene glycol (PEG) precipitation; (c) resuspension in Tween 80/Tris solution and chloroform extraction; (d) the second PEG precipitation; (e) viral RNA extraction with TRIzol and isopropanol precipitation; and (f) RT-PCR combined with semi-nested PCR. The overall recovery of elution/concentration was 19.5% with poliovirus. The whole procedure usually takes 19 hr. The overall detection sensitivity was 4 RT-PCR units of genogroup I norovirus (NoV) and 6.4 RT-PCR units of genogroup II Nov/25 g of oysters initially seeded. The virus-detecting method developed in this study should facilitate the detection of low levels of NoV in oysters.

Microarray Study of Genes Differentially Modulated in Response to Nitric Oxide in Macrophages

  • Nan, Xuehua;Maeng, Oky;Shin, Hyo-Jung;An, Hyun-Jung;Yeom, Young-Il;Lee, Hay-Young;Paik, Sang-Gi
    • Animal cells and systems
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    • v.12 no.1
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    • pp.15-21
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    • 2008
  • Nitric oxide(NO) has been known to play important roles in numerous physiologic processes including neurotransmission, vasorelaxation, and cellular apoptosis. Using a mouse cDNA gene chip, we examined expression patterns and time course of NO-dependent genes in mouse macrophage RAW264.7 cells. Genes shown to be upregulated more than two fold or at least at two serial time points were further selected and validated by RT-PCR. Finally, 81 selected genes were classified by function as signaling, apoptosis, inflammation, transcription, translation, ionic homeostasis and metabolism. Among those, genes related with signaling, apoptosis and inflammation, such as guanylate cyclase 1, soluble, alpha3(Gucy1a3); protein kinase C, alpha($Pkc{\alpha}$); lymphocyte protein tyrosine kinase(Lck); BCL2/adenovirus E1B 19 kDa-interacting protein(Bnip3); apoptotic protease activating factor 1(Apaf1); X-linked inhibitor of apoptosis(Xiap); cyclin G1(Ccng1); chemokine(C-C motif) ligand 4(Ccl4); B cell translocation gene 2, anti-proliferative(Btg2); lysozyme 2(Lyz2); secreted phosphoprotein 1(Spp1); heme oxygenase(decycling) 1(Hmox1); CD14 antigen(Cd14); and granulin(Grn) may play important roles in NO-dependent responses in murine macrophages.

Characterization and Functional Study of PyrR Orthologues from Genome Sequences of Bacteria (세균 게놈 유래성 PyrR Orthologue의 기능 분석)

  • 김사열;조현수;설경조;박승환
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.103-110
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    • 2003
  • The regulation of pyrimidine nucleotide synthesis has been proved to be controlled by a regulatory protein PyrR-mediated attenuation in the Gram-positive bacteria. After several bacterial genome sequencing projects, we have discovered the PyrR orthologues in the databases for Haemophilus influenzae and Synechocystis and sp. PCC6803 genome sequences. To investigate whether these PyrR orthologue proteins regulate pyrimidine nucleotide synthesis as well as the cases of Bacillus, the PyrR regions of each strains were amplified by PCR and cloned with pUC19 or T-vector in Escherichia coli and with a shuttle vector pHPS9 for E. coli and B. subtilis. For the regulation test of the PyrR orthologues, the aspartate-transcarbamylase (ATCase) assay was carried out. From the results of the ATCase assay, it was confirmed that Synechocystis sp. PCC6803 could not restore by pyrimidines to a B. subtilis, PyrR but H. influenzae PyrR could. For Purification of PyrR orthologue proteins, PyrR orthologue genes were cloned into the expression vector (pET14b). Over-expressed product of PyrR orthologue genes was purified and analyzed by the SDS-PACE. The purified PyrR orthologue proteins from H. influenzae and Synechocystis sp. PCC6803 turned out to be molecular mass of 18 kDa and 21 kDa, respectively. The result of uracil phosphoribosyl transferase (UPRTase) assay with purified PyrR orthologue proteins showed that H. influenzae PyrR protein only has UPRTase activity. In addition, we could predict several regulatory mechanisms that PyrR orthologue proteins regulate pyrimidine de novo synthesis in bacteria, through phylogenetic analysis for PyrR orthologue protein sequences.

The Effects of Hand-Acupuncture Therapy on Headaches in Children (고려수지요법이 아동의 두통에 미치는 효과)

  • Hong Yeon-Ran
    • Child Health Nursing Research
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    • v.11 no.4
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    • pp.427-435
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    • 2005
  • Purpose: The purpose of this study was to identify the effects of hand acupuncture therapy on headaches in children. Method: A quasi experimental pre-test and post (1,2,3)-test design was used. Data were collected from March 2 to April 16 2001. Forty children were assigned to an experimental (20) or control group (20). The experimental group received Hand-Acupuncture therapy on the meridian point; A30, A31, A32, A33, E8, I2, M2, M3, M5, B25, B26, B27 for each 3 minutes, while the control group rested on a bed. Data were analyzed using the SAS program with $x^2$-test, t-test, repeated measured ANOVA, and ANCOVA. Result: In the experimental group, descriptive headache intensity (f=64.33, p=0.00), numeric headache intensity (F=74.69, 122.50, 7.52. p=0.00), and medication requirements ($x^2$=19.00, p=0.00) were significantly lower than those of the control group. Conclusion: These findings indicate that hand acupuncture therapy is effective for reduction of headaches. Therefore, hand acupuncture therapy can be considered an independent nursing intervention for reducing headaches in children.

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Dependence of Mouse Embryonic Development in vitro on the Exposed Period to Oviductal Environment (난관체류시간에 따른 생쥐초기배의 체외발생능력)

  • Song, H.B.;Seo, B.B.;Kim, K.S.;Park, S.E.;Lee, S.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.19 no.2
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    • pp.117-123
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    • 1992
  • Development in vitro of 2-cell mouse embryos was examined after appropriate exposure to oviductal milieu to demonstrate biological activity present in the oviducts. ICR and ($C57Bl/6{\times}Balb/c$) $F_1$ hybrid mice were superovulated and mated for the recovery of early embryos. Embryos were recoverd at every 2h intervals from 32h post-hCG(hph) to 56 hph. The proportions of developmental stages were determined in the recovered embryos. Development in vitro of 2-cell embryos was more rapid in $F_1$ hybrid than in ICR, showing high proportions of 4-cell embryo and blastocyst at 120 hph. 100% of blastocyst development was obtained at 38hph in $F_1$ hybrid and at 50 hph in ICR when 2-cell embryos were cultured upto 120hph in vitro. Moreover, in vitro culture of oviducts containing 2-cell embryos in ICR mice for 12h from 34hph to 46hph increased developmental capacity of ICR mouse embryo in vitro. The results indicate that oviductal environment contains substances having mitogenic activity and overcoming early cell block in vitro. The mitogenic activity is effective in vitro as well as in vivo.

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