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Purification and Characterization of an Alkaliphilic Alginate Lyase AlgMytC from Saccharophagus sp. Myt-1

  • Sakatoku, Akihiro;Tanaka, Daisuke;Nakamura, Shogo
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.872-877
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    • 2013
  • In a previous study, we isolated and reported a second species of the Saccharophagus genus, Saccharophagus sp. strain Myt-1. In the present study, an alginate lyase gene (algMytC) from the genomic DNA of Myt-1 was cloned and characterized. The DNA sequence fragment obtained contained an open reading frame of 1,032 bp that encoded a protein of 343 amino acids with an estimated molecular mass of 37.6 kDa and a pI of 6.60. The deduced protein, AlgMytC, had the conserved amino acid sequences (RTELREM, QIH, YFKAGVYNQ) of the polysaccharide lyase family 7. A BLAST homology search indicated that AlgMytC shared an amino acid sequence identity of 95.9% with alg7A of S. degradans 2-40. The cloned and purified AlgMytC protein showed optimal activity at $40^{\circ}C$, and retained more than 90% of its total activity even after treatment at $25^{\circ}C$ for 24 h. AlgMytC was very alkaliphilic with an optimal pH of 9.0, and more than 90% of its activity was retained in the pH range 8.5-10.0. Moreover, AlgMytC was stable over a wide pH range. The activity of AlgMytC was also stable in the presence of various detergents.

Identification of differentially expressed Genes by methyl mercury in neuroblastoma cell line using SSH

  • Kim, Youn-Jung;Chang, Suk-Tai;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2002.10a
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    • pp.167-167
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    • 2002
  • Methylmercury (MeHg), one of the heavy metal compound, can cause severe damage to the central nervous system in humans. Many reports have contributed MeHg poisoning to contaminated foods and release into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established. To find genes differentially expressed by MeHg in neuronal cell, we peformed forward and reverse suppression subtractive hybridization (SSH) method on mRNA derived from neuroblastoma cell line, SH-SY5Y treated with solvent (DMSO) and 6.25 uM (IC$\sub$50/) MeHg. Differentially expressed CDNA clones were sequenced and the mRNAs were re-examined on Northern blots. These sequences were identified by BLAST homology search to known genes or expressed sequence tags (ESTs). Analysis of these sequences has provided an insight into the biological effects of MeHg in the pathogenesis of neurodegenerative disease and a possibility to develop more efficient and exact monitoring system of heavy metals as common environmental pollutants.

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C-Ring Cleavage of Isoflavones Daidzein and Genistein by a Newly-Isolated Human Intestinal Bacterium Eubacterium ramulus Julong 601

  • Wang, Xiu-Ling;Kim, Ki-Tae;Lee, Je-Hyeon;Hur, Hor-Gil;Kim, Su-Il
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.766-771
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    • 2004
  • Julong 601, a Gram-positive anaerobic bacterium strain capable of cleaving the C-ring of isoflavones daidzein and genistein, was isolated from human feces. BLAST search revealed that its complete 16S rDNA gene sequence has 99% similarity to Eubacterium ramulus. Metabolites of daidzein and genistein were determined as O-desmethylangolensin (O-Dma) and 2-(4-hydroxyphenyl) propionic acid (2-HPPA), respectively, based on UV, EI-MS, and $^1H$ and ^{13}C$ NMR spectral analyses. Enantiomers of O-Dma and 2-HPPA were isolated by chiral stationary-phase HPLC (CSP HPLC). Cleavage of the C-ring of daidzein and genistein by strain Julong 601 was highly enantioselective. Specific rotation ([$\alpha]_D$) and circular dichroism (CD) spectra of the enantiomers are reported here for the first time. Biotransformation kinetics of daidzein and genistein indicated that the C-ring of genistein has a higher susceptibility to bacterial degradation than that of daidzein.

Cloning and characterization of a gene encoding ABP57, a soluble auxin-binding protein

  • Lee, Keunpyo;Kim, Myung-Il;Kwon, Yu-Jihn;Kim, Minkyun;Kim, Yong-Sam;Kim, Donghern
    • Plant Biotechnology Reports
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    • v.3 no.4
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    • pp.293-299
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    • 2009
  • Auxin-binding protein 57 ($ABP_{57}$), a soluble auxin-binding protein, acts as a receptor to activate plasma membrane (PM) $H^+-ATPase$. Here, we report the cloning of abp57 and the biochemical characterization of its protein expressed in E. coli. The analysis of internal amino acid sequences of $ABP_{57}$ purified from rice shoots enabled us to search for the corresponding gene in protein DB of NCBI. Further BLAST analysis showed that rice has four abp57-like genes and maize has at least one homolog. Interestingly, Arabidopsis seems to have no homolog. Recombinant $ABP_{57}$ expressed in E. coli caused the activation of PM $H^+-ATPase$ regardless of the existence of IAA. Scatchard analysis showed that the recombinant protein has relatively low affinity to IAA as compared to natural $ABP_{57}$. These results collectively support the notion that the cloned gene is responsible for $ABP_{57}$.

Human RPS4X/Y Genes and Pseudogene Family: Chromosomal Localization and Phylogenetic Analysis

  • Lee, Ji-Won;Yi, Joo-Mi;Shin, Kyung-Mi;Kim, Heui-Soo
    • Journal of Life Science
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    • v.11 no.2
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    • pp.81-82
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    • 2001
  • The human ribosomal protein 54 genes, RPS4X and RPS4Y are located on the X and Y chromosomes. They have been postulated as candidate for Turner syndrome which was characterized by gonadal dysgenesis, short stature, and various external and internal anomalies. Using the BLAST search program, we identified sixteen RPS4 pseudogenes from the human genome and analyzed them phylogenetically. The RPS4-C12-1, C12-2, and C12-3 pseudogenes from chromosome 12 have been evolved independently during hominid evolution. The RPS4X gene from X chromosome it closely related to the RPS4-C12-2 from chromosome 12 and RPS4-C5 from chromosome 5, whereas the RPS4Y gene is very closely related to RPS4-C16 from chromosome 16. The exact mapping of the RPS4 pseudogene family was peformed, indicating that the RPS4 pseudogene family was mapped on human chromosomes 1, 2, 5, 6, 8, 10, 11, 12, 13, 16, 18, 19 and 20. Taken together, the precise chromosomal localization and phylegenetic relationship of the RPS4 pseudo-genes could be of great use in further study for understanding the Turner syndrome.

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Isolation and characterization of unrecorded yeasts species in the family Metschnikowiaceae and Bulleribasidiaceae in Korea

  • Park, Yuna;Maeng, Soohyun;Srinivasan, Sathiyaraj
    • Journal of Species Research
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    • v.9 no.3
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    • pp.198-203
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    • 2020
  • The goal of this study was to isolate and identify wild yeasts from soil samples. The 15 wild yeast strains were isolated from the soil samples collected in Pocheon city, Gyeonggi Province, Korea. Among them, four yeast stains were unrecorded, and 11 yeast stains were previously recorded in Korea. To identify wild yeasts, microbiological characteristics were observed by API 20C AUX kit. Pairwise sequence comparisons of the D1/D2 domain of the 26S rRNA were performed using Basic Local Alignment Search Tool(BLAST). Cell morphology of yeast strains was examined by phase contrast microscope. All strains were oval-shaped and polar budding and positive for assimilation of glucose, 2-keto-ᴅ-gluconate, N-acetyl-ᴅ-glucosamine, ᴅ-maltose and ᴅ-saccharose (sucrose). There is no official report that describes these four yeast species: one strain of the genus Kodamaea in the family Metschnikowiaceae and three strains of the Hannaella in the family Bulleribasidiaceae. Kodamaea ohmeri YI7, Hannaella kunmingensis YP355, Hannaella luteola YP230 and Hannaella oryzae YP366 were recorded in Korea, for the first time.

A Development of Auto-lnterlock Relation Generating System for Electronic Interlocking Equipment (전자연동장치를 위한 연동데이터 자동 생성 시스템의 개발)

  • Kwon, Cheol;Lee, Ki-Chul;Choi, Sung-Bum;Lee, Jin-Ha
    • Proceedings of the KSR Conference
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    • 2009.05a
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    • pp.831-839
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    • 2009
  • An interlocking equipment of railway signalling systems is manufactured with electrical devices and electrical interlocking equipment has been substituted for existing interlock equipment(relay sequential logic type). But interlock conditions are still generated from rail diagram and used to make Interlock Table manually. In order to make EIS(Electrical Interlock System) operate, we should write interlock data which is made from interlock table. But, as the station is larger and more complicated, handwork may becomes a very tediou work and makes more mistakes. Therefore the development of CAD system for Interlocking System is very significant, if CAD can reduce the mistakes from handwork and help the configuring the interlocking system. In this paper, we first arrange some rules which can be used to extract route information automatically from rail diagram and interlocking rules. And then we propose "Search-And-Rollback" algorithm to extract route information and individual interlocking rules. The proposed algorithm is implemented and tested through the signal design process of the Hyundai-Steel private railway to carry melted pig iron from the blast furnace to the steel-making workshop. some cases. It shows that CAD for Interlocking system is very helpful in time saving aspect and system reliability.

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Geographic Variation of Granulilittorina exigua (Littorinidae, Gastropoda) in Korea Based on the Mitochondrial Cytochrome b Gene Sequence

  • Song, Jun-Im;Suh, Jae-Hwa;Kim, Sook-Jung
    • Animal cells and systems
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    • v.4 no.3
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    • pp.267-272
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    • 2000
  • Partial sequence of the mitochondrial cytochrome b gene was analyzed to investigate genetic variation from 10 geographic populations of Granulilittorina exigua in Korea. The sequence of 282 base pairs was determined by PCR-directed silver sequencing method. The sequences of two species within the genus Littorina reserved in NIH blast search were utilized to determine geographic variations of species referred. The levels of mtDNA sequence differences were 0.00-2.54% within populations and 0.71-4.43% between populations. There were four amino acid differences between representative species of the genera Granulilittorina and Littorina, but no differences within populations of the genus Granulilittorina. The UPGMA and the N-J trees based on Tamura-Nei genetic distance matrix were constructed, which showed that the genus Granulilittorina was divided into three groups such as eastern (even exception for Tokdo population), southern, and western regional populations. The degrees of genetic divergence within populations of each group were p=0.021, p=0.019, and p=0.018, respectively. The divergence between the eastern and southern populations was p=0.032, showing closer relationship than with the western populations (p=0.052). Based on the diverged time estimation, the eastern and southern populations of Granulilittorina exigua in Korea diverged from the western populations about 2.1 MYBP, and the eastern and southern populations diverged from each other about 1.3 MYBP.

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Design and Implementation of a PCR Primer Search System on Cloud Computing Environments (클라우드 컴퓨팅 환경에서 PCR Primer 검색 시스템 설계 및 개발)

  • Park, Junho;Lim, Jongtae;Kim, Dongjoo;Lee, Yunjeong;Ryu, Eunkyung;Ahn, Minje;Cha, Jaehong;Yu, Seok Jong;Yoo, Jaesoo
    • Proceedings of the Korea Contents Association Conference
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    • 2012.05a
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    • pp.269-270
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    • 2012
  • 유전자 증폭을 위한 정확한 PCR Primer의 디자인은 핵심적인 기반 기술이다. 기존 연구를 통해 각 유전자별 특이적인 PCR Primer를 디자인할 수 있는 도구가 제안되었으나, 유전체 정보를 활용한 대단위의 디자인작업을 수행하기에는 적합하지 않았다. 본 논문에서는 클라우드 컴퓨팅 환경에서 대규모의 유전체를 대상으로 특이적인 PCR Primer를 디자인하고 검색할 수 있는 시스템을 설계하고 구현한다. 제안하는 시스템은 Hadoop 플랫폼에서의 MapReduce 프레임워크를 기반으로 설계 및 구현하여 유전자 서열검색을 대규모로 수행할 수 있도록 하였다. 5만개의 질의를 이용한 성능 평가 결과, 제안하는 기법은 기존 BLAST를 이용한 검색방법에 비해 약 3배의 성능 향상을 보였다.

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Solution State Structure of pB1, the Mimotopic Peptide of Apolipoprotein B-100, by NMR

  • Lee, Sung-Ran;Kim, Dae-Sung;Kim, Hyo-Joon;Lee, Yong-Woo;Won, Ho-Shik
    • Bulletin of the Korean Chemical Society
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    • v.25 no.12
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    • pp.1845-1849
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    • 2004
  • Apolipoprotein B-100 (Apo-B100) is a major protein component for low density lipoproteins (LDL). A number of mimetic peptides of Apo-B100 were screened from the phase-displayed random peptide library by utilizing monoclonal antibody (B9). Mimetic peptide for B9 epitope against apo B-100 was CRNVPPIFNDVYWIAF (pB1). From the BLAST search, the mimetic peptide pB1 had 40% homology with apo B-100. As a result of the structural determination of this mimotope using homo/hetero nuclear 2D-NMR techniques and NMR-based distance geometry (DG)/molecular dynamic (MD) computations, DG structure had low penalty value of 0.3-0.6 ${\AA}^2$ and the total RMSD was 0.5-1.5 ${\AA}. Moreover, pB1 structure included a weak $3_{10}$-helix from $Ile^7$,/TEX> to $Trp^{13}$.