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Cloning, Expression, and Characterization of a Cold-Adapted Lipase Gene from an Antarctic Deep-Sea Psychrotrophic Bacterium, Psychrobacter sp. 7195

  • Zhang, Jinwei;Lin, Shu;Zeng, Runying
    • Journal of Microbiology and Biotechnology
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    • 제17권4호
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    • pp.604-610
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    • 2007
  • A psychrotrophic strain 7195 showing extracellular lipolytic activity towards tributyrin was isolated from deep-sea sediment of Prydz Bay and identified as a Psychrobacter species. By screening a genomic DNA library of Psychrobacter sp. 7195, an open reading frame of 954 bp coding for a lipase gene, lipA1, was identified, cloned, and sequenced. The deduced LipA1 consisted of 317 amino acids with a molecular mass of 35,210 kDa. It had one consensus motif, G-N-S-M-G (GXSXG), containing the putative active-site serine, which was conserved in other cold-adapted lipolytic enzymes. The recombinant LipA1 was purified by column chromatography with DEAE Sepharose CL-4B, and Sephadex G-75, and preparative polyacrylamide gel electrophoresis, in sequence. The purified enzyme showed highest activity at $30^{\circ}C$, and was unstable at temperatures higher than $30^{\circ}C$, indicating that it was a typical cold-adapted enzyme. The optimal pH for activity was 9.0, and the enzyme was stable between pH 7.0-10.0 after 24h incubation at $4^{\circ}C$. The addition of $Ca^{2+}\;and\;Mg^{2+}$ enhanced the enzyme activity of LipA1, whereas the $Cd^{2+},\;Zn^{2+},\;CO^{2+},\;Fe^{3+},\;Hg^{2+},\;Fe^{2+},\;Rb^{2+}$, and EDTA strongly inhibited the activity. The LipA1 was activated by various detergents, such as Triton X-100, Tween 80, Tween 40, Span 60, Span 40, CHAPS, and SDS, and showed better resistance towards them. Substrate specificity analysis showed that there was a preference for trimyristin and p-nitrophenyl myristate $(C_{14}\;acyl\; groups)$.

기관리포지터리 수용모형 연구: 과학분야 연구자를 중심으로 (The Adoption Model of Institutional Repositories: Which Constructs Attract Scientists to Share Their Research Outputs?)

  • 황혜경;이지연
    • 정보관리학회지
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    • 제34권2호
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    • pp.47-80
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    • 2017
  • 본 연구의 목적은 과학분야 연구자를 대상으로 기관리포지터리 수용에 영향을 미치는 요인들을 도출하고 이들 요인들간의 관계를 규명함으로써 기관리포지터리 수용모형을 개발하는 것이다. 270명의 물리수학분야와 생명과학분야 연구자들이 응답한 설문조사와 12명의 심층면담 내용 분석 결과, 개인의 심리적 특성이 기관리포지터리 수용의도에 가장 높은 영향을 미치는 것으로 나타났다. 그 다음은 성과기대, 사회조직적 영향 순으로 확인되었으나, 인지된 위험은 유의한 영향을 주지 못하는 것으로 검증되었다. 기관리포지터리 수용의도 향상을 위한 전략으로 국가주도적 제도 개선 방안과 운영 기관의 역할강화 및 홍보 방안, 그리고 연구자의 참여유도형 서비스방안을 제언하였다.

Development of a Virtual Simulator for Agile Manufacturing System

  • C., Sangmin;C., Younghee;B., Jongil;L., Manhyung
    • 제어로봇시스템학회:학술대회논문집
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    • 제어로봇시스템학회 2000년도 제15차 학술회의논문집
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    • pp.103-103
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    • 2000
  • In this paper to cope with the reduction of products life-cycle as the variety of products along with the various demands of consumers, a virtual simulator is developed to make the changeover of manufacturing line efficient to embody a virtual simulation similar to a real manufacturing line. The developed virtual simulator can design a layout of a factory and make the time scheduling. Every factory has one simulator so that one product can be manufactured in the factories to use them as virtual factories. We suggest a scheme that heightens the agility to the diversity of manufacturing models by making the information of manufacturing lines and products models to be shared. The developed unit simulator can construct a proper virtual manufacturing line along with the required process of products using several kinds of operator and work cell. A user with the simulator can utilize an interface that makes one to manage the separate task process for each manu(acturing module, change operator components and work cells, and easily teach tasks of each task module. The developed simulator was made for users convenience by Microsoft Visual C++ 6.0 that can develop a program supplying graphic user interface environment and by OpenGL of the Silicon Graphics as a graphic library to embody 3D graphic environment. Also, we show that the simulator can be used efficiently for the agile manufacturing by the communication among the factories being linked by TCP/IP and a hybrid database system made by a hierarchical model and a relational model being developed to standardize the data information.

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Characterization and Induction of Potato HMGR genes in Relation to Antimicrobial Isoprenoid Synthesis

  • Park, Doil;Richard M. Bostock
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 1995년도 Proceedings of special lectures on Molecular Biological Approaches to Plant Disease National Agricultural Science and Technology Institute Suwon, Korea
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    • pp.55-75
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    • 1995
  • Induction of HMG-Co A reductase (HMGR) is essential for the biosynthesis of sesquiterpenoid phytoalexins and steroid derivatives in Solanaceous plants following wounding and pathogen infection. To better understand this complex step in stress-responsive isoprenoid synthesis, three classes of cDNAs for HMGR (hmg1, hmg2, and hmg3) were isolated from a potato tuber library. The potato cDNAs had extensive homology in open reading frames but had low homology in the 3'-untranslated regions. RNA gel blot analysis using gene-specific probes revealed that hmg1 is induced by wounding but wound induction is strongly suppressed by arachidonic acid or by inoculation with Phytophthora infestants. In contrast, hmg2 and hmg3 are slightly induced by wounding and strongly enhanced by arachidonic acid or inoculation. The induction and suppression of HMGR genes parallel the suppression of steroid and stimulation of sesquiterpenoid accumulations observed in earlier investigations. Treatment of the tuber disks with a low concentration of methyl-jasmonate doubled the wound induced accumulation of hmg1 transcripts and steroid-glycoalkaloid accumulation, but did not affect the abundance of transcripts for hmg2 or hmg3 nor induce phytoalexins. High concentration of methyl-jasmonate suppressed hmg1 mRNA and steroid-glycoalkaloid accumulation, induced hmg3 mRNA, and did not elicit phytoalexins. Lipoxygenase inhibitors suppressed the accumulation of of hmg1 transcripts and steroid-glycoalkaloids, which were restored by exogeneous methyl-jasmonate. Methyl-jasmonate applied together with arachidonic acid enhanced the elicitor induced accumulation of sesquiterpenes and sustained steroid-glycoalkaloid levels with transcript levels for the various HMGR mRNAs equal to or greater than wound-only treatment. These results domonstrate that the consequences of wound- and pathogen-responses of plants are different at the levels of gene expression and associated secondary metabolism.

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High Spatial Resolution Satellite Image Simulation Based on 3D Data and Existing Images

  • La, Phu Hien;Jeon, Min Cheol;Eo, Yang Dam;Nguyen, Quang Minh;Lee, Mi Hee;Pyeon, Mu Wook
    • 한국측량학회지
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    • 제34권2호
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    • pp.121-132
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    • 2016
  • This study proposes an approach for simulating high spatial resolution satellite images acquired under arbitrary sun-sensor geometry using existing images and 3D (three-dimensional) data. First, satellite images, having significant differences in spectral regions compared with those in the simulated image were transformed to the same spectral regions as those in simulated image by using the UPDM (Universal Pattern Decomposition Method). Simultaneously, shadows cast by buildings or high features under the new sun position were modeled. Then, pixels that changed from shadow into non-shadow areas and vice versa were simulated on the basis of existing images. Finally, buildings that were viewed under the new sensor position were modeled on the basis of open library-based 3D reconstruction program. An experiment was conducted to simulate WV-3 (WorldView-3) images acquired under two different sun-sensor geometries based on a Pleiades 1A image, an additional WV-3 image, a Landsat image, and 3D building models. The results show that the shapes of the buildings were modeled effectively, although some problems were noted in the simulation of pixels changing from shadows cast by buildings into non-shadow. Additionally, the mean reflectance of the simulated image was quite similar to that of actual images in vegetation and water areas. However, significant gaps between the mean reflectance of simulated and actual images in soil and road areas were noted, which could be attributed to differences in the moisture content.

Cloning and Expression of a Rice cDNA Encoding a Lls1 Homologue of Maize

  • Jwa, Nam-Soo;Park, Sang-Geun;Park, Chan-Ho;Kim, Soon-Ok;Ahn, Il-Pyung;Park, Sook-Young;Yoon, Choong-Hyo;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • 제16권3호
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    • pp.151-155
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    • 2000
  • A cDNA (Oslls1) encoding Lls1-homologue of maize was isolated from cDNA library of rice (Oryza sativa cv. Ilpum). The 2,138 bp of full length Oslls1 clone contains an open reading frame of 1,623 nucleotides encoding 575 amino acid residues. The deduced amino acid sequence of Oslls1 has a high level of homology with chlorophyll a oxygenases of Arabidopsis thaliana (67%) and Marchantia polymorpha (65%). Southern blot analysis of genomic DNA indicates the existence of a small gene family for Oslls1 in the rice genome. The expression of Oslls1 mRNA was induced in leaves and germinating seeds. Treatment of $H_2O$$_2$significantly down-regulated Oslls1 expression. The expression of Oslls1 mRNA was consititutively down-regulated in the blm, a rice mutant exhibiting spontaneous necrotic lesions. These results suggest that this Oslls1 gene may be involved incell death mechanisms in the blm mutant of rice.

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Molecular Cloning of the Sec61p ${\gamma}$ Subunit Homologue Gene from the Mole Cricket, Gryllotalpa orientalis

  • Kim, Iksoo;Lee, Kwang-Sik;Jin, Byung-Rae;Kim, Eun-Sun;Lee, Heui-Sam;Ahn, Mi-Young;Sohn, Hung-Dae;Ryu, Kang-Sun
    • International Journal of Industrial Entomology and Biomaterials
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    • 제5권1호
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    • pp.73-77
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    • 2002
  • The Sec61 trimeric complex ($\alpha$,$\beta$, and ${\gamma}$ subunits) is one of the Sec-complex responsible for post-translational protein translocation across the endoplasmic reticulum membrane in diverse organisms. In this study, a cDNA encoding the Sec61p ${\gamma}$ subunit homologue was isolated from the cDNA library of the mole cricket, Gryllotalpa orientalis. Sequence analysis of a 442-bp cDNA clone showed it to contain an open reading frame of 68 amino acid residues consisted of 204-bp. The homologues of the gene were found in the GenBank database in a diverse organism including insect, mammals, fungi, and plants. The deduced amino acid sequence of Sec61p ${\gamma}$ subunit homologue of the mole cricket showed the highest homology to the gene of the singly known insect, Drosophila melanogester (93% identity), and the least homology to that of the baker's yeast, Saccharomyces cerevisiae (37.2%). Phylogenetic analysis also confirmed a close relationship between the insect Sec61p ${\gamma}$ subunit homologues of G. orientalis and D. melanogester. Hydropathy analysis of the cricket mole and published other data suggested that the hydrophobic segment close to C-terminus is predicted to be the putative membrane anchor, Multiple alignment of the Sec61p ${\gamma}$ subunit homologue among several organisms showed the presence of several conserved domains including the conserved proline at position 28.

Molecular cloning and characterization of an antigenic protein with a repeating region from clonorchis sinensis

  • Kim, Tae-Yun;Kang, Shin-Yong;Ahn, Il-Young;Cho, Seung-Yull;Hong, Sung-Jong
    • Parasites, Hosts and Diseases
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    • 제39권1호
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    • pp.57-66
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    • 2001
  • In the course of immunoscreening of Clonorchis sinensis cDNA library, a cDNA CsRP12 containing a tandem repeat was isolated. The cDNA CsRP 12 encodes two putative peptides of open reading frames (ORFs) 1 and 2 (CsRP12-1 and -2). The repetitive region is composed of 15 repeats of 10 amino acids. Of the two putative peptides, CsRP12-1 was proline-rich and found to have homologues in several organisms. Recombinant proteins of the putative peptides were bacterially produced and purified by an affinity chromatography Recombinant CsRP12-1 protein was recognized by sera of clonorchiasis patients and experimental rabbits, but recombinant CsRP 12-2 was not. One of the putative peptide, CsRP12-1, is designated CsPRA, proline-rich antigen of C. sinensis. Both the C-termini of CsRP12-1 and -2 were bacterially produced and analysed to show no antigenicity. Recombinant CsPRA protein showed high sensitivity and specificity. In experimental rabbits, IgG antibodies to CsPRA was produced between 4 and 8 weeks after the infection and decreased thereafter over one you. These results indicate that CsPRA is equivalent to a natural protein and a useful antigenic protein for serodiagnosis of human clonorchiasis.

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New Tumor Metastasis Suppressor Gene from Korean Tiger Shark (Scyliorhinus torazame)

  • CHO Jung Jong;LEE Jae Hyung;LEE Sang-Jun;LIM Woon Ki;KIM Yung-Jin;KIM Kyu-Won;KIM Young Tae
    • 한국수산과학회지
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    • 제30권6호
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    • pp.984-991
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    • 1997
  • New tumor suppressor gene, snm23, homologous to human nm23/NDP kinase (human nucleoside diphosphate kinase) gene whose product has a tumor metastasis inhibitory activity, was first cloned from Korean tiger shark (Scyliorhinus forazame) skin cDNA library constructed by using a $\lambda$ ZAP-II cDNA synthesis kit. About $1\times10^5$ plaques were screened and several positive plaques were isolated and confirmed by second screening. The phagemid containing a positive clone from the Uni-Zap XR vector was excised in vivo and the gene containing the tumor metastasis suppressor protein was named as snm23. Cloned gene, snm23, was sequenced with ABI-PRISM 310 Genetic Analyzer. The nucleotide and deduced amino acid sequences of snm23 have shown an open reading frame consisting of 450 base pairs that correspond to a protein of 150 amino acid residues, with a calculated molecular mass of 16.8 kDa. Sequence comparison of snm23 with human nm23/NDP kinase was performed by using Blast protein data base of National Center for Biotechnology Information. In order to determine tissue specificity, reverse transcription-polymerase chain reaction (RT-PCR) was used. Good expression level of snm23/NDP kinase was detected at the tissues from skin, cartilage, and liver of Korean tiger shark.

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Methylovorus sp. Strain SS1 DSM11726으로부터 rpoH 유전자의 클로닝과 염기서열 분석 (Cloning and Nucleotide Sequence Analysis of the rpoH Gene from Methylovorus sp. Strain SS1 DSM11726)

  • 엄치용;송승은;박미화;김영민
    • 한국미생물·생명공학회지
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    • 제35권3호
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    • pp.177-183
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    • 2007
  • 열충격 시그마인자를 코딩하는 유전자 rpoH가 결여된 돌연변이체 대장균(Escherichia coli satrain A7448)을, 메탄올 자화세균인 Methylovorus sp. strain SS1 DSM11726의 phagemid library로 형질전환 시켜서 $30^{\circ}C$에서 성장하는 Escherichia coli strain A7448 로부터 Methylovorus sp. strain SS1 DSM11726의 rpoH 유전자를 클로닝하고 그 염기서열을 분석하였다. 1,793-bp 염기서열 분석 결과 Methylovorus sp. strain SS1 DSM11726의 RpoH는 284개의 아미노산으로 이루어져 있었으며 예상된 분자량은 32,006, p1값은 5.79로 나타났으며, 동일계열의 ${\beta}$-proteobacteria에 속하는 세균들의 RpoH와 높은 상동성을 보여주었다. Methylovorus sp. strain SS1 DSM11726의 RpoH는 대장균의 RpoH의 기능을 대신할 수 있음을 보여주었다. 열충격 후 RpoH양은 15분까지 지속적으로 증가하다 20분 뒤 양이 감소하는 양상을 나타내었다. 이는 Methylovorus sp. strain SS1 DSM11726의 RpoH 단백질 역시 열에 의해 유도됨을 말해 준다.