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Identifikasi dan Kloning Gen CalBsyn ke dalam Plasmid pJ912-AGα-RFP di Escherichia coli
No full textSistem tampilan permukaan khamir telah menjadi teknik yang semakin
popular untuk rekayasa protein dan pembuatan pustaka protein. Protein heterolog
yang ditampilkan pada permukaan sel khamir diekspresikan dalam bentuk fusi
dengan suatu protein penahan atau protein permukaan. Protein CALB (Candida
antratica Lipase B) yang disandikan oleh gen sintetik CalBsyn dapat ditingkatkan
efektivitasnya melalui ekspresi pada permukaan sel khamir. Plasmid pJ912-AGα-
RFP mengandung gen penyandi protein permukaan, AGα (Agglutinin-α) dan gen
penyandi protein penanda ekspresi, RFP (Red fluorescent protein). Penelitian ini
bertujuan untuk memperoleh konstruksi vektor pJ912-AGα-RFP-CalBsyn untuk
ekspresi protein CALB rekombinan pada permukaan sel khamir Pichia pastoris
melalui kloning gen CalBsyn ke dalam plasmid pJ912-AGα-RFP pada Escherichia
coli. Gen CalBsyn tanpa sinyal sekresi natifnya diperoleh dari plasmid pJ912-
CALB melalui amplifikasi PCR menggunakan pasangan primer mCalB-Sal-F dan
mCalB-His-Sal-R. Gen mCalBsyn yang diperoleh berukuran panjang 1004 bp. Baik
gen mCalBsyn maupun plasmid pJ912-AGα-RFP dipotong dengan enzim restriksi
SalI, kemudian diligasi dan ditransformasi ke dalam E. coli dengan metode kejut
panas. Transformasi menghasilkan 11 koloni transforman putatif yang selanjutnya
dipurifikasi menjadi koloni-koloni tunggal. Analisis PCR dengan primer AOX1-F
dan mCalB-His-Sal-R menunjukkan adanya 34 koloni tunggal dari klon yang
dianalisis terindikasi positif membawa plasmid rekombinan pJ912-AGα-RFPCalBsyn
Rancang Bangun vacuum Penyedot debu Sebagai fasilitas galangan Mini Jurusan teknik Perkapalan politeknik Negeri Bengkalis
Get PDFBengkalis State Polytechnic Mini Shipbuilding Engineering Shipyard often encounters work in the process of building fiberglass ships. Every time you carry out the manufacturing process, of course you first make a mold and when making the mold you first cut or carve the part of the plywood or board that you want to make a mold from. After carrying out the sanding or cutting process, of course the dust from the fibers is scattered on the floor. Usually this cleaning is
done using a broom because the vacuum cleaner in the yard is unstable and therefore cannot be used anymore. The dust in the workshop is dust that comes from wood fibers and fibres. In simple terms, the working principle of a vacuum cleaner is a suction and exhaust process using a number of physical principles to clean dirt effectively. Tool design is the process of creating a drawing using the AutoCAD application, where at this stage the author will design the shape of the
vacuum cleaner. As for how the vacuum cleaner works. What the author made is not much different from the vacuum cleaner sold in stores, it's just that the materials needed are different
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
Bioassay of Recombinant Human Granulocyte Colony Stimulating Factor (rhG-CSF) for Neutropenia Treatment in Male Sprague Dawley Rats
Get PDFBackground: Recombinant human granulocyte colony stimulating factor (rhG-CSF) is a first line therapy for neutropenia. However, it is less affordable for most patients in developing and poor countries. Therefore, biosimilar products are developed to suppress the cost of treatment, namely with rhG-CSF. This study aimed to explore the establishment of an affordable rhG-CSF that has similar potential to induce neutrophils recovery as the positive control.Materials and Methods: The rhG-CSF was expressed as inclusion body in Escherichia coli NiCo21(DE3). The inclusion body was then solubilized, refolded, purified and characterized prior to be used in the bioactivity assay. Cyclophosphamide-induced male Sprague Dawley rats were used as animal model and administered with rhG-CSF. Blood sample was collected at several points of time, before and after rhG-CSF treatments. Complete blood count and peripheral blood smear were conducted to investigate the activity of the rhG-CSF on each blood cells type, particularly neutrophil.Results: Specific activity on neutrophil proliferation was shown after treatments with our rhG-CSF and positive control. Positive control dose 40 mg/kg BW was statistically similar with that of the rhG-CSF dose 80 and 120 mg/kg BW. However, in neutropenic condition, recovery of neutrophil counts could not be achieved within 4 days of treatments. Thus, a longer treatment is needed to observe the activity of the rhG-CSF as an antineutropenia agent.Conclusion: The rhG-CSF has been proven having specific activity on neutrophil proliferation. However, improvement in the rhG-CSF preparation is still needed and longer administration of the rhG-CSF has to be applied in the future study.Keywords: rhG-CSF, biosimilar, neutropenia, Sprague Dawley rat
Konstruksi Gen Glukosa Oksidase untuk Subkloning ke dalam Plasmid pPICZα dengan Penambahan Oligonukleotida Penyandi Spacer Peptida
No full textGlukosa oksidase (EC 1.1.3.4) merupakan flavoenzim yang berasal dari
Aspergillus niger. Enzim Glukosa Oksidase (GOX) intraseluler yang berasal dari
A. niger memiliki aktivitas yang lebih tinggi dibandingkan dengan aktivitas enzim
GOX ekstraselulernya. Enzim GOX intraseluler yang berasal dari A. niger memiliki
tingkat produksi yang rendah serta proses pemurnian yang cukup rumit. Penelitian
ini bertujuan untuk mendapatkan gen penyandi GOX yang siap untuk disisipkan ke
dalam plasmid pPICZα untuk kemudian diekspresikan pada Pichia pastoris secara
ekstraseluler. Perbanyakan plasmid rekombinan pGEM®-T Easy-GOX dilakukan
pada sel bakteri Escherichia coli DH5α. Konstruksi gen GOX dilakukan dengan
amplifikasi menggunakan PCR. Amplifikasi dilakukan sebanyak dua kali. Ukuran
panjang basa yang diperoleh dari hasil amplifikasi kedua sebesar 1787 bp, terdiri
atas situs restriksi XhoI dan oligonukleotida penyandi spacer peptida pada ujung 5’
sekuens gen GOX, serta situs restriksi XbaI pada ujung 3’ sekuens gen GOX
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