• Title/Summary/Keyword: Partial Encoding

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Molecular Characterization of a Protein Kinase Gene in Chiness Cabbage(Brassica campestrics subsp. napus var. pekinensis)

  • Jeong, Sang-Ho;Ahn, Ji-Hoon;Lee, June-Seung;Lee, Jong-Seob
    • Animal cells and systems
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    • v.1 no.1
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    • pp.135-142
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    • 1997
  • Random sequencing of expressed sequence tags in roots of Chinese cabbage led to isolation of a partial cDNA clone, BR77, which encoded a putative protein kinase. Using the BR77 cDNA as a probe, we isolated a full-length cDNA encoding the Brassica campestris protein kinase 1 (Bcpk1). The Bcpt1 cDNA contained one open reading frame encoding a polypeptide of 439 amino acids. The putative polypeptide consisted of a short N-terminal region and a protein kinase catalytic domain. The catalytic domain of Bcpkl showed a high homology to cAMP- and calcium- phospholipid-dependent subfamilies of serine/threonine protein kineses. Eleven major catalytic domains in protein kineses were well conserved in Bcpk1. However, Bcpk1 contained a unique nonhomologous intervening sequence between subdomains VII and VIII, which was not found in protein kineses of animals and lower eukaryotes. Genomic DNA gel blot analysis showed that Bcpt1 genes might be present as three copies in the Chinese cabbage genome. These imply that Bcpk1 belongs to a plant-specific serine/threonine protein kinase subfamily.

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Spontaneous Release of Bacteriophage Particles by Lactobacillus rhamnosus Pen

  • Jarocki, Piotr;Podlesny, Marcin;Pawelec, Jaroslaw;Malinowska, Agata;Kowalczyk, Sylwia;Targonski, Zdzislaw
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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    • pp.357-363
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    • 2013
  • The identification of bacteriophage proteins on the surface of Lactobacillus rhamnosus Pen was performed by LC-MS/MS analysis. Among the identified proteins, we found a phage-derived major tail protein, two major head proteins, a portal protein, and a host specificity protein. Electron microscopy of a cell surface extract revealed the presence of phage particles in the analyzed samples. The partial sequence of genes encoding the major tail protein for all tested L. rhamnosus strains was determined with specific primers designed in this study. Next, RT-PCR analysis allowed detection of the expression of the major tail protein gene in L. rhamnosus strain Pen at all stages of bacterial growth. The transcription of genes encoding the major tail protein was also proved for other L. rhamnosus strains used in this study. The present work demonstrates the spontanous release of prophage-encoded particles by a commercial probiotic L. rhamnosus strain, which did not significantly affect the bacterial growth of the analyzed strain.

Characterization of a Soil Metagenome-Derived Gene Encoding Wax Ester Synthase

  • Kim, Nam Hee;Park, Ji-Hye;Chung, Eunsook;So, Hyun-Ah;Lee, Myung Hwan;Kim, Jin-Cheol;Hwang, Eul Chul;Lee, Seon-Woo
    • Journal of Microbiology and Biotechnology
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    • v.26 no.2
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    • pp.248-254
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    • 2016
  • A soil metagenome contains the genomes of all microbes included in a soil sample, including those that cannot be cultured. In this study, soil metagenome libraries were searched for microbial genes exhibiting lipolytic activity and those involved in potential lipid metabolism that could yield valuable products in microorganisms. One of the subclones derived from the original fosmid clone, pELP120, was selected for further analysis. A subclone spanning a 3.3 kb DNA fragment was found to encode for lipase/esterase and contained an additional partial open reading frame encoding a wax ester synthase (WES) motif. Consequently, both pELP120 and the full length of the gene potentially encoding WES were sequenced. To determine if the wes gene encoded a functioning WES protein that produced wax esters, gas chromatography-mass spectroscopy was conducted using ethyl acetate extract from an Escherichia coli strain that expressed the wes gene and was grown with hexadecanol. The ethyl acetate extract from this E. coli strain did indeed produce wax ester compounds of various carbon-chain lengths. DNA sequence analysis of the full-length gene revealed that the gene cluster may be derived from a member of Proteobacteria, whereas the clone does not contain any clear phylogenetic markers. These results suggest that the wes gene discovered in this study encodes a functional protein in E. coli and produces wax esters through a heterologous expression system.

A Queriable XML Compression using Inferred Data Types (추론한 데이타 타입을 이용한 질의 가능 XML 압축)

  • ;;Chung Chin-Wan
    • Journal of KIISE:Databases
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    • v.32 no.4
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    • pp.441-451
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    • 2005
  • HTML is mostly stored in native file systems instead of specialized repositories such as a database. Like HTML, XML, the standard for the exchange and the representation of data in the Internet, is mostly resident on native file systems. However. since XML data is irregular and verbose, the disk space and the network bandwidth are wasted compared to those of regularly structured data. To overcome this inefficiency of XML data, the research on the compression of XML data has been conducted. Among recently proposed XML compression techniques, some techniques do not support querying compressed data, while other techniques which support querying compressed data blindly encode data values using predefined encoding methods without considering the types of data values which necessitates partial decompression for processing range queries. As a result, the query performance on compressed XML data is degraded. Thus, this research proposes an XML compression technique which supports direct and efficient evaluations of queries on compressed XML data. This XML compression technique adopts an encoding method, called dictionary encoding, to encode each tag of XML data and applies proper encoding methods for encoding data values according to the inferred types of data values. Also, through the implementation and the performance evaluation of the XML compression technique proposed in this research, it is shown that the implemented XML compressor efficiently compresses real-life XML data lets and achieves significant improvements on query performance for compressed XML data.

Cloning and Expression of the Cathepsin F-like Cysteine Protease Gene in Escherichia coli and Its Characterization

  • Joo, Han-Seung;Koo, Kwang-Bon;Park, Kyun-In;Bae, Song-Hwan;Yun, Jong-Won;Chang, Chung-Soon;Choi, Jang-Won
    • Journal of Microbiology
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    • v.45 no.2
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    • pp.158-167
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    • 2007
  • In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.

Multiplex RT-PCR Assay for the Detection of Apple stem grooving virus and Apple chlorotic leaf spot virus in Infected Korean Apple Cultivars

  • Park, Hong-Lyeol;Yoon, Jae-Seung;Kim, Hyun-Ran;Baek, Kwang-Hee
    • The Plant Pathology Journal
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    • v.22 no.2
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    • pp.168-173
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    • 2006
  • To develop the diagnostic method for the viral infection in apple, the partial genes corresponding to the N-terminal region of RNA polymerase of Apple stem grooving virus (ASGV) and coat protein of Apple chlorotic leaf spot virus (ACLSV) were characterized from the infected apple cultivars in Korea. Based on the nucleotide sequences of the characterized partial genes, the virus gene-specific primers were designed for the detection of ASGV and ACLSV infected in species of Malus. The RT-PCR using the primers for the genes of ASGV and ACLSV successfully gave rise to 404 and 566 bp DNA fragments, respectively. Using those viral gene-specific primers, the multiplex RT-PCR assays were also established to diagnose the mixed infection by ASGV and ACLSV simultaneously. Furthermore, the control primers, which have to be included for the RT-PCR as an internal control, were designed using the nucleotide sequence of the gene encoding elongation factor $1{\alpha}(EF1{\alpha})$. This multiplex RT-PCR including the control primers provides more reliable, rapid and sensitive assay for the detection of ASGV and ACLSV infected in Korean apple cultivars.

Cloning and Sequencing of Gene Fragment of Acid Proteinase from Penicillium oxalicum HCLF-34 (Penicillium oxalicum HCLF-34로부터 Acid Proteinase의 부분유전자 Cloning 및 Sequencing)

  • 현성희;천재순;강상순;김진규
    • Korean Journal of Microbiology
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    • v.40 no.1
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    • pp.12-16
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    • 2004
  • Acid proteinase has been discovered in Aspergillus niger (acid protease A) and Cryphonectria parasitica (acid proteinase EapC) and it plays major roles in cheese formation from milk. In this study, a partial gene encoding acid proteinase in Penicillium oxalicum HCLF-34 was cloned by using PCR with degenerate primers corresponding to highly conserved regions of the acid proteinase. The partial acid proteinase gene in P. oxalicum HCLF-34 contains an open reading frame of 438 base pairs and encodes an acid proteinase protein of 146 amino aicds. The predicted amino acid sequences showed 71 % homology with acid protease A and 67% homology with EapC.

A Efficient Architecture of MBA-based Parallel MAC for High-Speed Digital Signal Processing (고속 디지털 신호처리를 위한 MBA기반 병렬 MAC의 효율적인 구조)

  • 서영호;김동욱
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.41 no.7
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    • pp.53-61
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    • 2004
  • In this paper, we proposed a new architecture of MAC(Multiplier-Accumulator) to operate high-speed multiplication-accumulation. We used the MBA(Modified radix-4 Booth Algorithm) which is based on the 1's complement number system, and CSA(Carry Save Adder) for addition of the partial products. During the addition of the partial product, the signed numbers with the 1's complement type after Booth encoding are converted in the 2's complement signed number in the CSA tree. Since 2-bit CLA(Carry Look-ahead Adder) was used in adding the lower bits of the partial product, the input bit width of the final adder and whole delay of the critical path were reduced. The proposed MAC was applied into the DWT(Discrete Wavelet Transform) filtering operation for JPEG2000, and it showed the possibility for the practical application. Finally we identified the improved performance according to the comparison with the previous architecture in the aspect of hardware resource and delay.

A New Genetic Algorithm for Shortest Path Routing Problem (최단 경로 라우팅을 위한 새로운 유전자 알고리즘)

  • ;R.S. Ramakrishna
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.27 no.12C
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    • pp.1215-1227
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    • 2002
  • This paper presents a genetic algorithmic approach to shortest path (SP) routing problem. Variable-length chromosomes (strings) and their genes (parameters) have been used for encoding the problem. The crossover operation that exchanges partial chromosomes (partial-routes) at positionally independent crossing sites and the mutation operation maintain the genetic diversity of the population. The proposed algorithm can cure all the infeasible chromosomes with a simple repair function. Crossover and mutation together provide a search capability that results in improved quality of solution and enhanced rate of convergence. Computer simulations show that the proposed algorithm exhibits a much better quality of solution (route optimality) and a much higher rate of convergence than other algorithms. The results are relatively independent of problem types (network sizes and topologies) for almost all source-destination pairs.

Characterization and Expression Pattern of the Partial Myostatin cDNA in Shrimp, Fenneropenaeus chinensis

  • Lee, Sang Beum;Kim, Yong Soo;Yoon, Moongeun;Kim, Su-Kyoung;Jang, In Kwon;Lim, Hyun Jeong;Jin, Hyung-Joo
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.4
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    • pp.224-229
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    • 2007
  • Muscle tissue expresses many muscle-specific genes, including myostatin (also known as GDF8) that is a member of the transforming growth factor-beta superfamily. Myostatin (MSTN) negatively regulates mammalian skeletal muscle growth and development by inhibiting myoblast proliferation. Mice and cattle possessing mutant MSTN alleles display a 'double muscling' phenotype characterized by extreme skeletal muscle hypertrophy and/or hyperplasia. In this study, we first have characterized partial cDNA of a MSTN gene from the muscle tissue in the F. chinensis and examined its expression pattern in various tissues. The partial MSTN gene (GenBank accession number EU 131093) in the F. chinensis was 1134 bp, encoding for 377 amino acids that showed 63-93% amino acid similarity to other vertebrate MSTNs, containing a conserved proteolytic cleavage site (RXRR) and conserved cysteine residues in the C-terminus. Based on a RT-PCR, the MSTN gene was expressed in the all tissues of F. chinensis used in this study.

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