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Transcription analysis and cloning of u32.002 collagenolytic peptidase from geobacillus lituanicus dsm 15325t.
No full textCollagens are one of the most important proteins including their functions and biotechnological application. The biotechnological application of collagens wouldn‘t be possible without the process of collagenolysis which is catalysed by collagenolytic peptidases. The aim of this work was to perform transcriptional analysis of U32.002 (Helicobacter-type) collagenolytic peptidase gene and its cloning as well as basic analysis. The analysis of U32.002 peptidase gene transcription was performed after the extraction of total RNA from G. lituanicus DSM 15325T cells that were in two different stages of growth. Reverse transcription assays were performed after total RNA extraction. After the process of reverse transcription samples of cDNA were used for the diagnostic PGR that was carried out by using five different primers. This gene was cloned with and without putative signal/pro sequence in order to produce the preparation of U32.002 peptidase. Full sequence of U32.002 peptidase gene was cloned into pTZ57R/T and then into the expression vector pET28c(+). U32.002 gene without putative signal/pro sequence was cloned into pJET1.2, and then into pET28c(+). SDS-PAGE and MALDI-TOF analyses were performed in order to determine the fact of expression in E. coli BL21 (DE3). Full scale expression optimisation was performed, as well. The influence of calcium and zinc ions to the structural stability of U32.002 peptidase with putative signal/pro sequence was analysed. The collagenolytic activity of U32.002 peptidase was analysed by using zymography of native collagen, type I. The U32.002 peptidase of G. lituanicus DSM 15325T is transcribed constitutively during the exponential phase of growth, whereas only fragments of U32.002 gene transcript are observed in the later growth phases. It is evident that zinc ions increase structural thermostability of U32.002. The homodimer of U32.002 peptidase with putative signal/pro sequence was able to digest native type I collagen at 50 °C; pH 7,4
Peptidazių iš Geobacillus thermoleovorans DSM 15325 kolagenolizinio aktyvumo analizė: M3 proteolizinių fermentų šeimos M3B pošeimio oligopeptidazės ir U32 proteolizinių fermentų šeimos peptidazės charakterizavimas.
No full textThe full understanding of laws of nature cannot be accomplished without comprehensive characterisation of collagenolysis as collagen is a major structural protein in organisms. The diversity of collagenolytic peptidases from non-pathogenic bacteria remains only fragmentary characterized, whereas the characterization of the diversity of collagenolytic peptidases from eukaryotes and pathogenic bacteria are far more comprehensive. The determined constitutive production of collagenases by moderate thermophile Geobacillus thermoleovorans DSM 15325 confirms the ability of thermophiles to be ever ready for collagen uptake. The identification of constitutive collagenolytic peptidases variety produced by geobacilli lead to substantial development of collagen degradation model and confirms importance of collagenolytic potential for adaptive plasticity of bacteria. Determined M3 family M3B subfamily oligopeptidases, secreted by geobacilli, characteristics indicate oligopeptidases nutritional importance for bacteria during the adaption for environmental changes. Combination of oligopeptidases characteristics also outlines biotechnological applicability of this thermostable hydrolase. Determined U32 family peptidase from geobacilli characteristics ensure objective understanding of U32 family peptidases characteristics and functional importance. U32 family peptidases ability to specifically in vitro interact with dsRNA was never determined for collagenolytic peptidases previously
The collagenolytic activity analysis of Geobacillus thermoleovorans DSM 15325 peptidases: characterization of M3 proteolytic enzymes family M3B subfamily oligopeptidase and U32 proteolytic enzymes family peptidase.
No full textThe full understanding of laws of nature cannot be accomplished without comprehensive characterisation of collagenolysis as collagen is a major structural protein in organisms. The diversity of collagenolytic peptidases from non-pathogenic bacteria remains only fragmentary characterized, whereas the characterization of the diversity of collagenolytic peptidases from eukaryotes and pathogenic bacteria are far more comprehensive. The determined constitutive production of collagenases by moderate thermophile Geobacillus thermoleovorans DSM 15325 confirms the ability of thermophiles to be ever ready for collagen uptake. The identification of constitutive collagenolytic peptidases variety produced by geobacilli lead to substantial development of collagen degradation model and confirms importance of collagenolytic potential for adaptive plasticity of bacteria. Determined M3 family M3B subfamily oligopeptidases, secreted by geobacilli, characteristics indicate oligopeptidases nutritional importance for bacteria during the adaption for environmental changes. Combination of oligopeptidases characteristics also outlines biotechnological applicability of this thermostable hydrolase. Determined U32 family peptidase from geobacilli characteristics ensure objective understanding of U32 family peptidases characteristics and functional importance. U32 family peptidases ability to specifically in vitro interact with dsRNA was never determined for collagenolytic peptidases previously
Going Beyond Counting First Authors in Author Co-citation Analysis
Get PDFThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Variations on the Author
Get PDF“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Get PDFWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
Dispelling the Myths Behind First-author Citation Counts
Get PDFWe conducted a full-scale evaluative citation analysis study of scholars in the XML research field to explore just how different from each other author rankings resulting from different citation counting methods actually are, and to demonstrate the capability of emerging data and tools on the Web in supporting more realistic citation counting methods. Our results contest some common arguments for the continued
use of first-author citation counts in the evaluation of scholars, such as high correlations between author rankings by first-author citation counts and other citation
counting methods, and high costs of using more realistic citation counting methods that are not well-supported by the ISI databases. It is argued that increasingly available digital full text research papers make it possible for citation analysis studies to go beyond what the ISI databases have directly supported and to employ more
sophisticated methods
koamabayili/VECTRON-author-checklist: VECTRON author checklist
No full textWe have done our best to complete the author checklist relating to the use of animals in the hut study. Note that the objective for the hut study was to evaluate the IRS treatment applications for residual efficacy against Anopheles mosquitoes, including the local An. coluzzii mosquito population. Cows were only used to attract mosquitoes into the huts and no tests were carried out directly on the cows. The author checklist is intended for use with studies where experiments are carried out on animals, which is why we have had such difficulty in completing this for the hut study, as many of the questions do not relate to how the cows were used
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