• Title/Summary/Keyword: Protein sequence search

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A New Putative Chitinase from Reticulitermes speratus KMT001

  • Ham, Youngseok;Park, Han-Saem;Kim, Yeong-Suk;Kim, Tae-Jong
    • Journal of the Korean Wood Science and Technology
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    • v.47 no.3
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    • pp.371-380
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    • 2019
  • Termites are pests that cause serious economic and cultural damage by digesting wood cellulose. Termites are arthropods and have an epidermis surrounded by a chitin layer. To maintain a healthy epidermis, termites have chitinase (${\beta}$-1,4-poly-N-acetyl glucosamidinase, EC 3.2.1.14), an enzyme that hydrolyzes the ${\beta}$-1,4 bond of chitin. In this study, the amino acid sequence of the gene, which is presumed to be termite chitinolytic enzyme (NCBI accession no. KC477099), was obtained from a transcriptomic analysis of Reticulitermes speratus KMT001 in Bukhan Mountain, Korea. An NCBI protein BLAST search confirmed that the protein is a glycoside hydrolase family 18 (GH18). The highest homology value found was 47%, with a chitinase from Araneus ventricosus. Phylogenetic analysis indicated that the KC477099 protein has the same origins as those of arthropods but has a very low similarity with other arthropod chitinases, resulting in separation at an early stage of evolution. The KC477099 protein contains two conserved motifs, which encode the general enzymatic characteristics of the GH18 group. The amino acid sequences $Asp^{156}-Trp^{157}-Glu^{158}$, which play an important role in the enzymatic activity of the GH18 group, were also present. This study suggests that the termite KC477099 protein is a new type of chitinase, which is evolutionarily distant from other insect chitinases.

Studies on the Epitope of Neuronal Growth Inhibitory Factor (GIF) with Using of the Specific Antibody

  • Pang, Li-Yan;Ru, Bing-Gen
    • BMB Reports
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    • v.38 no.6
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    • pp.646-649
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    • 2005
  • Human neuronal growth inhibitory factor (GIF), a metalloprotein classified as metallothionein-3, is specifically expressed in mammal central nervous system (CNS). In these Studies the specific antibody to human GIF was prepared and used to search the epitope of human GIF by enzyme-linked immunosorbent assay (ELISA) and sequence comparison. The result of ELISA showed the epitope of human GIF may locate on a octapeptide (EAAEAEAE) in the $\alpha$-domain of human GIF, and the result of nerve cell culture indicated that the biological activity of GIF may be affected by the specific antibody.

Complete Sequence of a Gene Encoding KAR3-Related Kinesin-like Protein in Candida albicans

  • Kim Min-Kyoung;Lee Young Mi;Kim Wankee;Choi Wonja
    • Journal of Microbiology
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    • v.43 no.5
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    • pp.406-410
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    • 2005
  • In contrast to Saccharomyces cerevisiae, little is known about the kinesin-like protein (KLP) in Candida albicans. The motor domain of kinesin, or KLP, contains a subregion, which is well conserved from yeast to humans. A similarity search, with the murine ubiquitous kinesin heavy chain region as a query, revealed 6 contigs that contain putative KLPs in the genome of C. albicans. Of these, the length of an open reading (ORF) of 375 amino acids, temporarily designated CaKAR3, was noticeably short compared with the closely related S. cerevisiae KAR3 (ScKAR3) of 729 amino acids. This finding prompted us to isolate a ${\lambda}$ genomic clone containing the complete CaKAR3 ORF, and here the complete sequence of CaKAR3 is reported. CaKAR3 is a C-terminus motor protein, of 687 amino acids, encoded by a non-disrupting gene. When compared with ScKAR3, the amino terminal region of 112 amino acids was unique, with the middle part of the 306 amino acids exhibiting $25\%$ identity and $44\%$ similarity, while the remaining C-terminal motor domain exhibited $64\%$ identity and $78\%$ similarity, and have been submitted to GeneBank under the accession number AY182242.

Molecular Identification and Sequence Analysis of Coat Protein Gene of Ornithogalum mosaic virus Isolated from Iris Plant

  • Yoon, Hye-In;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • v.18 no.5
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    • pp.251-258
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    • 2002
  • A potyvirus was isolated from cultivated Iris plants showing leaf streak mosaic symptom. Reverse transcription and polymerase chain reaction (RT-PCR) product of 1 kb long which encoded partial nuclear inclusion B and N-terminal region of viral coat protein (CP) genes for potyviruses was successfully amplified with a set of potyvirus-specific degenerate primers with viral RNA samples from the infected leaves: The RT-PCR product was cloned into the plasmid vector and its nucleotide sequences were determined. The nucleotide sequence of a CDNA clone revealed that the virus was an isolate of Ornithogalum moseic virus (OrMV) based on BLAST search analysis and was denoted as OrMV Korean isolate (OrMV-Ky). To further characterize the CP gene of the virus, a pair of OrMV-specific primers was designed and used for amplification of the entire CP gene of OrMV-Kr, The virus was easily and reliably detected from virus-infected Iris leaves by using the RT-PCR with the set of virus-specific primers. The RT-PCR product of the CP gene of the virus was cloned and its sequences were determined from selected recombinant CDNA clones. Sequence analysis revealed that the CP of OrMV-Kr consisted of 762 nucleotides, which encoded 253 amino acid residues. The CP of OrMV-Ky has 94.1-98.0% amino acid sequence identities (20 amino acid alterations) with that of other three isolates of OrMV, Two NT rich potential N-glycosylation motif sequences, NCTS and NWTM, and a DAC triple box responsible for aphid transmission were conserved in CPs of all the strains of OrMV. The virus has 58.5-86.2% amino acid sequence identities with that of other 16 potyviruses, indicating OrMV to be a distinct species of the genus. OrMV-Ky was the most related with Pterostylia virus Yin the phylogenetic tree analysis of CP at the amino acid level. This is the first report on the occurrence of OrMV in Iris plants in Korea. Data in this study indicate that OrMV is found in cultivated Iris plants, and may have mixed infection of OrMV and Iris severe mosaic virus in Korea.

Construction of Large Library of Protein Fragments Using Inter Alpha-carbon Distance and Binet-Cauchy Distance (내부 알파탄소간 거리와 비네-코시 거리를 사용한 대규모 단백질 조각 라이브러리 구성)

  • Chi, Sang-mun
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.19 no.12
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    • pp.3011-3016
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    • 2015
  • Representing protein three-dimensional structure by concatenating a sequence of protein fragments gives an efficient application in analysis, modeling, search, and prediction of protein structures. This paper investigated the effective combination of distance measures, which can exploit large protein structure database, in order to construct a protein fragment library representing native protein structures accurately. Clustering method was used to construct a protein fragment library. Initial clustering stage used inter alpha-carbon distance having low time complexity, and cluster extension stage used the combination of inter alpha-carbon distance, Binet-Cauchy distance, and root mean square deviation. Protein fragment library was constructed by leveraging large protein structure database using the proposed combination of distance measures. This library gives low root mean square deviation in the experiments representing protein structures with protein fragments.

A Cloning of Novel Esterase from a Metagenomic Library

  • Yoon, Sang-Young;Kim, Seung-Bum;Ryu, Yeon-Woo
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.243-246
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    • 2005
  • A novel esterase showing high enantioselectivity to (S)-ketoprofen ethyl ester was selected from fosmid environmental DNA library which is provided by Microbial Genomic & Applications Center. As a result of Blast search, the gene wasn't registerated in Gene Bank yet. And as we know, conserved domain region of esterase , G-X-S-X-G, wasn't discovered.$^{4)}$ And it is similar to Beta-lactamase. The DNA sequence of cloned esterase include an open reading frame consisting of 1170 bp, designated as EST-Y29, encoding a protein of 389 amino acids with a molecular mass of about 42.8 kDa. And amino acid sequence analysis revealed only a few identity (28%) to tile known esterases/lipases in the databases containing the conserved sequence motifs of esterases/lipases. when being comparison to other esterase revealed , this enzyme seems to be classified as a new member of esterase family. EST-Y29 was functionally overexpressed in a soluble form in E. coli with maximum conversion yield of (S)-ketoprofen at $65^{\circ}C$. This study demonstrates that functional screening combined with the sequential uses of restriction enzymes to exclude already known enzymes is a useful approach for isolating novel enzyme from a metagenome.

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Unification System for Analysis of DNA Sequence (DNA 서열 분석을 위한 통합 시스템)

  • Song, Young-Ohk;Chang, Duk-Jin
    • The Journal of the Korea Contents Association
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    • v.11 no.3
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    • pp.65-72
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    • 2011
  • We stand at real world that some practical use method of gene information appears in succession by entrance on the stage of advanced techonlogy. As a lot of studies and development are achieved based on analysis of bio data, necessity of a tool that can help correct interpretation of data is required more and more in a lot of targets of bioinformatics to search new relation and information are established. In this paper, we are offered in existing I wish to offer user a more convenient study tool developing system that can supplement shortcomings of various tools for data analysis. So we've designed to offer in united environment that is not environment that is parted ORF driving out, bio information retrieval and work of similarity comparison lamp to work for bio data analysis and offers lacking consecutiveness in existing analysis system.

Characteristics of Structure and Expression Pattern of ADSF/resistin Gene in Korean Native Cattle

  • Kang, Hye Kyeong;Park, Ji Ae;Seo, Kang Seok;Kim, Sang Hoon;Choi, Yun Jai;Moon, Yang Soo
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.3
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    • pp.329-334
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    • 2006
  • Adipocyte-specific secretory factor (ADSF)/resistin, a hormone, is a small cysteine-rich protein secreted from adipose tissue and has been implicated in modulating adipogenesis in humans and rodents. The objective of this study was to clone a gene encoding ADSF/resistin and to characterize its function in Korean Native Cattle (Hanwoo). The coding sequence was 330 base pairs and it encoded a protein of 109 amino acids. An NCBI BLAST-search revealed the cloned cDNA fragment shared significant homology (82%) with the cDNA encoding the human ADSF/resistin. The nucleotide sequence homology of the Hanwoo sequence was 73% and 64% for the rat and mouse, respectively. A 654 bp ADSF/resistin gene promoter was cloned and putative binding sites of transcription factors were identified. Tissue distribution of ADSF mRNA was examined in liver, skeletal muscles (tenderloin, biceps femoris), subcutaneous fat, and perirenal fat by RT-PCR. ADSF mRNAs were detected in fat tissues but not in liver and muscles, suggesting that ADSF/resistin expression may be induced during adipogenesis. Although, the physiological function of ADSF/resistin in the cow remains to be determined, these data indicate ADSF is related to the adipocyte phenotype and may have a possibly regulatory role in adipocyte function.

Computational Analysis of the 3-D structure of Human GPR87 Protein: Implications for Structure-Based Drug Design

  • Rani, Mukta;Nischal, Anuradha;Sahoo, Ganesh Chandra;Khattri, Sanjay
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7473-7482
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    • 2013
  • The G-protein coupled receptor 87 (GPR87) is a recently discovered orphan GPCR which means that the search of their endogenous ligands has been a novel challenge. GPR87 has been shown to be overexpressed in squamous cell carcinomas (SCCs) or adenocarcinomas in lungs and bladder. The 3D structure of GPR87 was here modeled using two templates (2VT4 and 2ZIY) by a threading method. Functional assignment of GPR87 by SVM revealed that along with transporter activity, various novel functions were predicted. The 3D structure was further validated by comparison with structural features of the templates through Verify-3D, ProSA and ERRAT for determining correct stereochemical parameters. The resulting model was evaluated by Ramachandran plot and good 3D structure compatibility was evidenced by DOPE score. Molecular dynamics simulation and solvation of protein were studied through explicit spherical boundaries with a harmonic restraint membrane water system. A DRY-motif (Asp-Arg-Tyr sequence) was found at the end of transmembrane helix3, where GPCR binds and thus activation of signals is transduced. In a search for better inhibitors of GPR87, in silico modification of some substrate ligands was carried out to form polar interactions with Arg115 and Lys296. Thus, this study provides early insights into the structure of a major drug target for SCCs.

A novel WD40 protein, BnSWD1, is involved in salt stress in Brassica napus

  • Lee, Sang-Hun;Lee, Jun-Hee;Paek, Kyung-Hee;Kwon, Suk-Yoon;Cho, Hye-Sun;Kim, Shin-Je;Park, Jeong-Mee
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.165-172
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    • 2010
  • Genes that are expressed early in specific response to high salinity conditions were isolated from rapeseed plant (Brassica napus L.) using an mRNA differential display method. Five PCR fragments (DD1.5) were isolated that were induced by, but showed different response kinetics to, 200 mM NaCl. Nucleotide sequence analysis and homology search revealed that the deduced amino sequences of three of the five cDNA fragments showed considerable similarity to those of ${\beta}$-mannosidase (DD1), tomato Pti-6 proteins (DD5), and the tobacco harpin-induced protein hin1 (DD4), respectively. In contrast, the remaining clones, DD3 and DD2, did not correspond to any substantial existing annotation. Using the DD3 fragment as a probe, we isolated a full-length cDNA clone from the cDNA library, which we termed BnSWD1 (Brassica napus salt responsive WD40 1). The predicted amino-acid sequence of BnSWD1 contains eight WD40 repeats and is conserved in all eukaryotes. Notably, the BnSWD1 gene is expressed at high levels under salt-stress conditions. Furthermore, we found that BnSWD1 was upregulated after treatment with abscisic acid, salicylic acid, and methyl jasmonate. Our study suggests that BnSWD1, which is a novel WD40 repeat-containing protein, has a function in salt-stress responses in plants, possibly via abscisic acid-dependent and/or -independent signaling pathways.