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The Molecular Study of Phosphotyrosine Protein Phosphatase (PtpA) from Streptomyces coelicolor A(3)2 (방선균이 생산하는 인산화타이로신 단백질 포스파타아제의 분자생물학적 연구)

  • 최학선;신용국;김춘성;김시욱
    • Journal of Life Science
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    • v.12 no.1
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    • pp.113-119
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    • 2002
  • The cloning and expression of Phosphotyrosine Protein Phosphatase into E. coli provides important tools of understanding of its functions and signal transduction mechanisms. The abundant soluble protein of the Phosphotyrosine Protein Phosphatase A (PtpA) and the active site mutant PtpA(C9S) were produced using the expression vector pET26 in E. coli and pIJ6021 with the thiostrepton in S. lividans. The enzyme activity of both proteins extracted by Ni-NTA column had same results from the expression vector pET26 and pIJ6021. The enzyme activity of phosphatase was found in the protein of PtpA, but not in that of C9S. The western blot detected by penta His-tag antibody resulted in the inducer, thiostrepton was not a good trigger to induce a large amount of PtpA protein. The overexpression of both proteins had no significantly different effect on the A factor cascade related to the secondary metabolite and mycelium formation between PtpA and C9S. However, overproduction of PtpA protein using pIJ6021 in S. lividans brought about a dramatic decrease in the amount of phosphotyrosine proteins (p200, p90, and p65), but no significantly phenotypic variation in S. lividans. This indicates that PtpA has an important proteome role in signal transduction mechanism of producing massive amount of phosphotyrosine protein in Streptomyces sp.

A Survey on the Installation Status of Plant-label in Elementary Schools: Focusing on Gwangju·Jeollanam-do (초등학교에 설치된 식물이름표 현황 조사연구 - 광주·전남권을 중심으로 -)

  • Kim, Tae-Hyeon;Hong, Mun-Gi
    • Journal of Korean Elementary Science Education
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    • v.43 no.1
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    • pp.35-47
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    • 2024
  • Thirty-five elementary schools in Gwangju and Jeollanam-do were surveyed on the installation status of plant labels. As a result of a comprehensive evaluation of the installation and condition of the plant labels, 12 out of 35 elementary schools were evaluated as "excellent," 13 as "good," and 8 as "insufficient." In some cases, various types of plant labels were installed on various types of plants and were updated continuously, indicating that the school members recognize the plant labels as an important educational tool. Meanwhile, some schools did not have a single plant label. Although plant labels were installed, instances occurred where they were neglected without continuous maintenance and management. Some plant labels were produced and installed by specialized companies, leading to errors in content or inadequate post-installation management, thereby compromising the school landscape. Therefore, we propose an alternative-allowing schools to independently produce and install plant labels. This approach preserves the school's uniqueness, and the process of producing and installing sustainable plant labels becomes an active educational tool.

Formation of the $CoSi_{2}$ using Co/Zr Bilayer on the Amorphous and the Single Crystalline Si Substrates (단결정과 비정질 Si 기판에서 Co/Zr 이중층을 이용한 $CoSi_{2}$ 형성)

  • Kim, Dong-Wook;Jeon, Hyeong-Tag
    • Korean Journal of Materials Research
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    • v.8 no.7
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    • pp.621-627
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    • 1998
  • The formation of Co-silicide between Co/Zr bilayer on the amorphous and crystalline Si substrates has been investigated. The films of Zr(50$\AA$) and Co(l50$\AA$) were deposited with e-beam evaporation system and were heattreated with the rapid thermal annealing system at the temperatures between 50$0^{\circ}C$ and 80$0^{\circ}C$ with 10$0^{\circ}C$ increments for 30 seconds. The phase identification of Co-silicide was carried out by XRD and the chemical analysis was examined by AES and RBS. The interface morphologies of Co/Zr bilayer films were investigated by cross sectional TEM and HRTEM. $CoSi_2$ was formed epitaxially on the crystalline Si substrate above $700^{\circ}C$ while polycrystalline $CoSi_2$ was grown on the amorphous Si substrate. The formation temperature of Co-silicide on the amorphous Si substrate was about 100 C lower than that on the crystalline Si. The COzSi phase was not identified on the both Si substrates. The formation temperature of first phase of Co-silicide on ColZr bilayer was higher than that on Co mono layer. CoSizlayer formed on the amorphous Si substrate exhibits better uniformity compared to the CoSiz formed on the crystalline substrate. The sheet resistance of CoSiz layer on crystalline Si was lower than that on the amorphous Si at high temperatures.tures.

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Cell Migration and Wound Healing Activities of Recombinant Thymosin β-4 Expressed in Escherichia coli (재조합 Thymosin β-4의 세포이동능과 상처치유능)

  • Hong, Kyo-Chang;Choi, Yung Hyun;Kim, Gun-Do;Cha, Hee-Jae;Jeon, Sung-Jong;Nam, Soo-Wan
    • Journal of Life Science
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    • v.32 no.2
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    • pp.135-141
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    • 2022
  • Thymosin β-4 (TB4) is a small peptide composed of 43 amino acids. To obtain sufficient biologically active mouse TB4 economically, we cloned and overexpressed this gene in an Escherichia coli system. With the isopropyl β-D-1-thiogalactopyranoside induction of the E. coli transformant, TB4 fusion protein with intein- and chitin-binding domain was successfully expressed in the soluble fraction within the E. coli cell. The TB4-intein - chitin-binding domain fusion protein was purified from the soluble fraction of E. coli cell lysate. The affinity chromatography with chitin beads and dithiothreitol-mediated intein self-cleavage reaction releases the TB4 peptide into the stripping solution. Sodium dodecyl sulphate - polyacrylamide gel electrophoresis and Western blot analyses were used to confirm that the recombinant TB4 peptide was produced with the expected size of 5 kDa. We found that the recombinant TB4 stimulated cell migration in the transwell plate chamber assay. After 18 hr of the treatment of the recombinant TB4 with 1 ng/ml concentration, the migration of the HT1080 cell was increased by 20% compared with that of the chemically synthesized TB4. The recombinant TB4 was also observed to promote the healing of a wound area in C57BL/6 mice by as high as 35% compared with that of the chemically synthesized TB4. These results suggest that the recombinant TB4 has better biological activity for cell migration and wound healing than that of the chemically synthesized TB4 peptide.

A Folksonomy Ranking Framework: A Semantic Graph-based Approach (폭소노미 사이트를 위한 랭킹 프레임워크 설계: 시맨틱 그래프기반 접근)

  • Park, Hyun-Jung;Rho, Sang-Kyu
    • Asia pacific journal of information systems
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    • v.21 no.2
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    • pp.89-116
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    • 2011
  • In collaborative tagging systems such as Delicious.com and Flickr.com, users assign keywords or tags to their uploaded resources, such as bookmarks and pictures, for their future use or sharing purposes. The collection of resources and tags generated by a user is called a personomy, and the collection of all personomies constitutes the folksonomy. The most significant need of the folksonomy users Is to efficiently find useful resources or experts on specific topics. An excellent ranking algorithm would assign higher ranking to more useful resources or experts. What resources are considered useful In a folksonomic system? Does a standard superior to frequency or freshness exist? The resource recommended by more users with mere expertise should be worthy of attention. This ranking paradigm can be implemented through a graph-based ranking algorithm. Two well-known representatives of such a paradigm are Page Rank by Google and HITS(Hypertext Induced Topic Selection) by Kleinberg. Both Page Rank and HITS assign a higher evaluation score to pages linked to more higher-scored pages. HITS differs from PageRank in that it utilizes two kinds of scores: authority and hub scores. The ranking objects of these pages are limited to Web pages, whereas the ranking objects of a folksonomic system are somewhat heterogeneous(i.e., users, resources, and tags). Therefore, uniform application of the voting notion of PageRank and HITS based on the links to a folksonomy would be unreasonable, In a folksonomic system, each link corresponding to a property can have an opposite direction, depending on whether the property is an active or a passive voice. The current research stems from the Idea that a graph-based ranking algorithm could be applied to the folksonomic system using the concept of mutual Interactions between entitles, rather than the voting notion of PageRank or HITS. The concept of mutual interactions, proposed for ranking the Semantic Web resources, enables the calculation of importance scores of various resources unaffected by link directions. The weights of a property representing the mutual interaction between classes are assigned depending on the relative significance of the property to the resource importance of each class. This class-oriented approach is based on the fact that, in the Semantic Web, there are many heterogeneous classes; thus, applying a different appraisal standard for each class is more reasonable. This is similar to the evaluation method of humans, where different items are assigned specific weights, which are then summed up to determine the weighted average. We can check for missing properties more easily with this approach than with other predicate-oriented approaches. A user of a tagging system usually assigns more than one tags to the same resource, and there can be more than one tags with the same subjectivity and objectivity. In the case that many users assign similar tags to the same resource, grading the users differently depending on the assignment order becomes necessary. This idea comes from the studies in psychology wherein expertise involves the ability to select the most relevant information for achieving a goal. An expert should be someone who not only has a large collection of documents annotated with a particular tag, but also tends to add documents of high quality to his/her collections. Such documents are identified by the number, as well as the expertise, of users who have the same documents in their collections. In other words, there is a relationship of mutual reinforcement between the expertise of a user and the quality of a document. In addition, there is a need to rank entities related more closely to a certain entity. Considering the property of social media that ensures the popularity of a topic is temporary, recent data should have more weight than old data. We propose a comprehensive folksonomy ranking framework in which all these considerations are dealt with and that can be easily customized to each folksonomy site for ranking purposes. To examine the validity of our ranking algorithm and show the mechanism of adjusting property, time, and expertise weights, we first use a dataset designed for analyzing the effect of each ranking factor independently. We then show the ranking results of a real folksonomy site, with the ranking factors combined. Because the ground truth of a given dataset is not known when it comes to ranking, we inject simulated data whose ranking results can be predicted into the real dataset and compare the ranking results of our algorithm with that of a previous HITS-based algorithm. Our semantic ranking algorithm based on the concept of mutual interaction seems to be preferable to the HITS-based algorithm as a flexible folksonomy ranking framework. Some concrete points of difference are as follows. First, with the time concept applied to the property weights, our algorithm shows superior performance in lowering the scores of older data and raising the scores of newer data. Second, applying the time concept to the expertise weights, as well as to the property weights, our algorithm controls the conflicting influence of expertise weights and enhances overall consistency of time-valued ranking. The expertise weights of the previous study can act as an obstacle to the time-valued ranking because the number of followers increases as time goes on. Third, many new properties and classes can be included in our framework. The previous HITS-based algorithm, based on the voting notion, loses ground in the situation where the domain consists of more than two classes, or where other important properties, such as "sent through twitter" or "registered as a friend," are added to the domain. Forth, there is a big difference in the calculation time and memory use between the two kinds of algorithms. While the matrix multiplication of two matrices, has to be executed twice for the previous HITS-based algorithm, this is unnecessary with our algorithm. In our ranking framework, various folksonomy ranking policies can be expressed with the ranking factors combined and our approach can work, even if the folksonomy site is not implemented with Semantic Web languages. Above all, the time weight proposed in this paper will be applicable to various domains, including social media, where time value is considered important.