• 제목/요약/키워드: Vector Synthesis

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산업용 효모에서 Bacillus subtilis Endo-$\beta$-1,4-Glucanase의 생합성 및 분비 (Synthesis and Secretion of the Endo-$\beta$-l,4-Glucanase from Bacillus subtilis in Industrial Yeast Strain)

  • 박용준;이영호;백운화;강현삼
    • 한국미생물·생명공학회지
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    • 제19권4호
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    • pp.348-355
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    • 1991
  • Bacillus subtilis의 Beta-1,4-glucanase 유전자와 Saccharomyces cerevisiae의 alcohol dehydrogenase isoenzyme I 유전자 (ADHI)의 promoter와 mouse $\alpha$-amylase의 분비신호를 연결하여 분비형 플라스미드를 구성하였으며 이를 산업용 알콜생산 효모인 Saccharomyces cerevisiae 54에 도입하여 형질전환시켰다. 한편 $\beta$-glucanase 유전자의 발현정도를 증대시키기 위해 CYC1 유전자의 전사종결신호를 부가한 후 역시 Saccharomyces cerevisiae 54에 도입하였다. 형질전체들은 carboxymethyl cellulose가 함유된 평판배지에서 $\beta$-glucanase를 분비하고 있음을 확인할 수 있었다. 전사종결 신호가 부가된 경우엔 전체역가가 2배 정도 증가하였다. 형질전환체들이 세포밖으로 분비한 효소역가는 전체의 60 정도였다.

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High-throughput and low-area implementation of orthogonal matching pursuit algorithm for compressive sensing reconstruction

  • Nguyen, Vu Quan;Son, Woo Hyun;Parfieniuk, Marek;Trung, Luong Tran Nhat;Park, Sang Yoon
    • ETRI Journal
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    • 제42권3호
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    • pp.376-387
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    • 2020
  • Massive computation of the reconstruction algorithm for compressive sensing (CS) has been a major concern for its real-time application. In this paper, we propose a novel high-speed architecture for the orthogonal matching pursuit (OMP) algorithm, which is the most frequently used to reconstruct compressively sensed signals. The proposed design offers a very high throughput and includes an innovative pipeline architecture and scheduling algorithm. Least-squares problem solving, which requires a huge amount of computations in the OMP, is implemented by using systolic arrays with four new processing elements. In addition, a distributed-arithmetic-based circuit for matrix multiplication is proposed to counterbalance the area overhead caused by the multi-stage pipelining. The results of logic synthesis show that the proposed design reconstructs signals nearly 19 times faster while occupying an only 1.06 times larger area than the existing designs for N = 256, M = 64, and m = 16, where N is the number of the original samples, M is the length of the measurement vector, and m is the sparsity level of the signal.

纖維質 資化性菌의 分子育種에 관한 硏究 -Cellulomonas속균의 ${\beta}$-glucosidase gene의 E. coli에의 cloning - (Studies on Molecular Improvement of Cellulose Utilizing Bacterial Strain -Molecular cloning of ${\beta}$-glucosidase gene of Cellulomonas sp. in E. coli-)

  • 배무;이재문
    • 미생물학회지
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    • 제22권3호
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    • pp.167-173
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    • 1984
  • The cellabiase (${\beta}$-glucosidase) gene in a Cellulomonas sp. CS1-1 was cloned into E. coli HB101 using the vector plasmid pBR322, and the expression of the gene in E. coli studied. The chromosomal DNA of the cellulomonas was digested by seveal restriction enzymes, each of which has only one cleaving site in plasmid pBR322. The recombinant plasmid, pSB2, created with Sal I frament, was expressed for the cellobiase gene in E. coli. The recombiant plasmid was estimated to contain 6.4 Kb foreign DNA at the Sal I site of plasmid pBR322 and the inserted DNA was mapped by single and double digestion with several enzymes. E. coli HB101(pSB2) has slowly grown in a mineral liquid medium containing cellobiose as a sole carbon source. The cellobiase activity in the transformed E. coli was 132 units per liter, which is equivalent to one twenty fifth of that in doner strain Cellulomonas sp. CS1-1. The transforned cell with plasmid containing cellulase gene grow well in the LB mediuns. The synthesis of cellobiase in the strain, E. coli HB101 (pSB2), was inhibited by glucose and at high concentration of cellobiose, and induced by cellobiose at low concentration.

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RNN을 이용한 Expressive Talking Head from Speech의 합성 (Synthesis of Expressive Talking Heads from Speech with Recurrent Neural Network)

  • 사쿠라이 류헤이;심바 타이키;야마조에 히로타케;이주호
    • 로봇학회논문지
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    • 제13권1호
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    • pp.16-25
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    • 2018
  • The talking head (TH) indicates an utterance face animation generated based on text and voice input. In this paper, we propose the generation method of TH with facial expression and intonation by speech input only. The problem of generating TH from speech can be regarded as a regression problem from the acoustic feature sequence to the facial code sequence which is a low dimensional vector representation that can efficiently encode and decode a face image. This regression was modeled by bidirectional RNN and trained by using SAVEE database of the front utterance face animation database as training data. The proposed method is able to generate TH with facial expression and intonation TH by using acoustic features such as MFCC, dynamic elements of MFCC, energy, and F0. According to the experiments, the configuration of the BLSTM layer of the first and second layers of bidirectional RNN was able to predict the face code best. For the evaluation, a questionnaire survey was conducted for 62 persons who watched TH animations, generated by the proposed method and the previous method. As a result, 77% of the respondents answered that the proposed method generated TH, which matches well with the speech.

ANALYSIS AND MANIPULATION OF CANDIDATE GENES FOR DIARRHEAL DISEASE VACCINE DEVELOPMENTS

  • Kim Young-Chang
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2000년도 International Meeting 2000
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    • pp.58-65
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    • 2000
  • Diarrheal diseases are a major cause of both illness and death in developing countries and are caused by rotavirus, Shigella spp., Salmonella spp., enterotoxigenic Escherichia coli (ETEC), and Vibrio spp. In this study, for the development of vaccine against diarrheal diseases caused by Shigella sonei, Salmonella typhimurium, E. coli O157, and Vibrio cholerae, cloning and nucleotide sequence analysis of genes and characteristics of their gene products in E. coli were performed. For construction of attenuated strain of S. sonnei KNIH104 and Salmonella typhimurium KNIH100, the aroA genes were cloned, respectively. The recombinant plasmid $_pJP{\Delta}A45$ containing aroA deleted region and suicide vector $(_pJP5603)$ was constructed. The aroA gene deleted mutants were constructed using this recombinant plasmid. For cloning gene encoding antigenic region of E. coli O157 KNIH317, the O-antigen synthesis gene cluster and sit gene was cloned. The E. coli XL1-Blue cells harboring this recombinant plasmid showed cytotoxicity in Vero cells. The ctx gene was cloned for tile purpose of antigenic region against V. cholerae KNIH002. Sequence analysis confirmed that the virulence gene cassette was consisted of ace, zot, ctxA and ctxB genes.

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Speaker-Dependent Emotion Recognition For Audio Document Indexing

  • Hung LE Xuan;QUENOT Georges;CASTELLI Eric
    • 대한전자공학회:학술대회논문집
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    • 대한전자공학회 2004년도 ICEIC The International Conference on Electronics Informations and Communications
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    • pp.92-96
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    • 2004
  • The researches of the emotions are currently great interest in speech processing as well as in human-machine interaction domain. In the recent years, more and more of researches relating to emotion synthesis or emotion recognition are developed for the different purposes. Each approach uses its methods and its various parameters measured on the speech signal. In this paper, we proposed using a short-time parameter: MFCC coefficients (Mel­Frequency Cepstrum Coefficients) and a simple but efficient classifying method: Vector Quantification (VQ) for speaker-dependent emotion recognition. Many other features: energy, pitch, zero crossing, phonetic rate, LPC... and their derivatives are also tested and combined with MFCC coefficients in order to find the best combination. The other models: GMM and HMM (Discrete and Continuous Hidden Markov Model) are studied as well in the hope that the usage of continuous distribution and the temporal behaviour of this set of features will improve the quality of emotion recognition. The maximum accuracy recognizing five different emotions exceeds $88\%$ by using only MFCC coefficients with VQ model. This is a simple but efficient approach, the result is even much better than those obtained with the same database in human evaluation by listening and judging without returning permission nor comparison between sentences [8]; And this result is positively comparable with the other approaches.

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The 3-[3α(2α-Hydroxy)pinane]-4,5-(pinan)-1,3-oxazolidine Synthesis, Structure and Properties

  • Bialek, Magdalena;Trzesowska, Agata;Kruszynski, Rafal
    • Bulletin of the Korean Chemical Society
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    • 제28권1호
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    • pp.89-94
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    • 2007
  • The new pinane derivative containing unique multifused ring system was synthesized. The crystal, molecular and electronic structure of the title compound has been determined. Both pinane ring systems have the same conformation. The five-membered oxazolidine ring exists in twisted chair conformation. The structure is expanded through O-H…O hydrogen bond to semiinfinite hydrogen-bonded chain. The bond lengths and angles in the optimised structure are similar to the experimental ones. The CH3 and CH2 groups (except this of oxazolidine ring) are negatively charged whereas the CH groups are positively charged. The largest negative potential is on the oxygen atoms. The C-N natural bond orbitals are polarised towards the nitrogen atom (ca. 61% at N) whereas the C-O bond orbitals are polarised towards the oxygen atom (ca. 67% at O). It is consistent with the charges on the nitrogen and oxygen atom of oxazolidine ring and the direction of the dipole moment vector (3.08 Debye).

Functional Implications in Apoptosis by Interferon Inducible Gene Product 1-8D, the Binding Protein to Adenovirus Preterminal Protein

  • Joung, In-Sil;Angeletti, Peter C.;Engler, Jeffrey A.
    • Journal of Microbiology
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    • 제41권4호
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    • pp.295-299
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    • 2003
  • Adenovirus (Ad) precursor to the terminal protein (pTP) plays an essential roles in the viral DNA replication. Ad pTP serves as a primer for the synthesis of a new DNA strand during the initiation step of replication. In addition, Ad pTP forms organized spherical replication foci on the nuclear matrix (NM) and anchors the viral genome to the NM. Here we identified the interferon inducible gene product 1-8D (Inid) as a pTP binding protein by using a two-hybrid screen of a HeLa cDNA library. Of the clones obtained in this assay, nine were identical to the Inid, a 13-kDa polypeptide that shares homology with genes 1-8U and Leu-13/9-27, most of which have little known functions. The entire open reading frame (ORF) of Inid was cloned into the tetracycline inducible expression vector in order to determine the biological functions related with adenoviral infection. When Inid was introduced to the cells along with adenoviruses, fifty to sixty percent of Ad-infected cells expressing Inid had rounded morphology, which was suggestive of apoptosis. Results from the terminal deoxynucleotidyl transferase (TdT) and DNA fragmentation assays confirmed that Inid induces apoptosis in Ad-infected or in uninfected cells. The Inid binding to pTP may target the cell for apoptotic destruction as a host defense mechanism against the viral infection.

Effect of Poly(ethylene glycol)Grafting on Polyethylenimine as a Gene Transfer Vector in vitro

  • 최진희;최준식;서혜란;박종상
    • Bulletin of the Korean Chemical Society
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    • 제22권1호
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    • pp.46-52
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    • 2001
  • To evaluate the non-ionic polymer, poly(ethylene glycol) (PEG), as a component in cationic copolymers for non-viral gene delivery systems, PEG was coupled to polyethylenimine (PEI). We present the effects of different degrees and shapes of pegylation of PEI on cytotoxicity, water solubility and transfection efficiency. This work reports the synthesis and characterization of a series of cationic copolymers on the basis of the conjugates of PEI with PEG. The modified molecules were significantly less toxic than the original polymer. Moreover, the chemical modification led to enhancement of their solubility. The comparison of pegylated PEIs with different degrees of derivation showed that all the polymers tested reached comparable levels of transgene expression to that of native PEI. As assessed by agarose gel electrophoresis, even highly substituted PEI derivatives were still able to form polyionic complexes with DNA. However, aside from an increase in solubility and retention of the ability to condense DNA, methoxy-PEG-modified PEIs resulted in a significant decrease in the transfection activity of the DNA complexes. In fact, the efficiency of the copolymer was compromised even at a low degree of modification suggesting that the PEG action resulting from its shape is important for efficient gene transfer. The mode of PEG grafting and the degree of modification influenced the transfection efficiency of PEI.

Isolation, Cloning and Co-Expression of Lipase and Foldase Genes of Burkholderia territorii GP3 from Mount Papandayan Soil

  • Putra, Ludwinardo;Natadiputri, Griselda Herman;Meryandini, Anja;Suwanto, Antonius
    • Journal of Microbiology and Biotechnology
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    • 제29권6호
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    • pp.944-951
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    • 2019
  • Lipases are industrial enzymes that catalyze both triglyceride hydrolysis and ester synthesis. The overexpression of lipase genes is considered one of the best approaches to increase the enzymatic production for industrial applications. Subfamily I.2. lipases require a chaperone or foldase in order to become a fully-activated enzyme. The goal of this research was to isolate, clone, and co-express genes that encode lipase and foldase from Burkholderia territorii GP3, a lipolytic bacterial isolate obtained from Mount Papandayan soil via growth on Soil Extract Rhodamine Agar. Genes that encode for lipase (lipBT) and foldase (lifBT) were successfully cloned from this isolate and co-expressed in the E. coli BL21 background. The highest expression was shown in E. coli BL21 (DE3) pLysS, using pET15b expression vector. LipBT was particulary unique as it showed highest activity with optimum temperature of $80^{\circ}C$ at pH 11.0. The optimum substrate for enzyme activity was $C_{10}$, which is highly stable in methanol solvent. The enzyme was strongly activated by $Ca^{2+}$, $Mg^{2+}$, and strongly inhibited by $Fe^{2+}$ and $Zn^{2+}$. In addition, the enzyme was stable and compatible in non-ionic surfactant, and was strongly incompatible in ionic surfactant.