• Title/Summary/Keyword: library characterization

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Characterization Strategies for Establishing City/County Records Centers in Kyonggi Region (특성화에 기반한 시.군 자료관 설립 방안 연구 - 경기도를 사례로 -)

  • Kim, Ik-Han
    • Journal of the Korean BIBLIA Society for library and Information Science
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    • v.15 no.1
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    • pp.155-175
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    • 2004
  • The records centers which belong to specific cities(-shi) or counties(-gun) are different in terms of constituencies and functions with the records centers of any other administrative agencies. The purpose of this study is to develop models of establishing the city/county records centers reflecting the recordkeeping environment of cities and counties especially located in Kyonggi Region. This present paper begins with analyzing the conditions of records management of cities and counties in Kyonggi Region, and the characterized functions of them. Based on these analyses, it suggests standardized procedures and characterization strategies for establishing city/county records centers.

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Screening and Characterization of an Esterase from a Metagenomic Library

  • KIM JEONG-NYEO;SEO MYUNG-JI;CHO EUN-AH;LEE SANG-JAE;KIM SEONG-BO;CHEIGH CHAN-ICK;PYUN YU-RYANG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1067-1072
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    • 2005
  • A metagenomic library was constructed using a fosmid vector, and total genomic DNA was extracted directly from soil at Cisolok (hot spring area, Indonesia). This library was composed of 10,214 clones and screened for lipolytic enzyme on tributyrin agar plates. An esterase gene (estMa) was subcloned and sequenced from a positive lipolytic active clone. Esterase EstMa was encoded by a 954-bp open reading frame and showed low ($11-33\%$) amino acid similarity to known esterases. The amino acid sequence analysis demonstrated that the enzyme is a new member of lipolytic enzyme family VI. The estMa gene encodes a preprotein of 317 amino acids with a predicted molecular mass of 34,799 Da. The purified enzyme exhibited optimal activity at $50^{\circ}C$ and pH 6.5. The $K_m,\;and\;V_{max}$ values of EstMa for the hydrolysis of p-nitrophenyl valerate were $45.3\;{\mu}M$ and 4.45 U/mg, respectively.

Korean BAC Library Construction and Characterization of HLA-DRA, HLA-DRB3

  • Park, Mi-Hyun;Lee, Hye-Ja;Bok, Jeong;Kim, Cheol-Hwan;Hong, Seong-Tshool;Park, Chan;Kimm, Ku-Chan;Oh, Berm-Seok;Lee, Jong-Young
    • BMB Reports
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    • v.39 no.4
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    • pp.418-425
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    • 2006
  • A human bacterial artificial chromosome (BAC) library was constructed with high molecular weight DNA extracted from the blood of a male Korean. This Korean BAC library contains 100,224 clones of insert size ranging from 70 to 150 kb, with an average size of 86 kb, corresponding to a 2.9-fold redundancy of the genome. The average insert size was determined from 288 randomly selected BAC clones that were well distributed among all the chromosomes. We developed a pooling system and three-step PCR screen for the Korean BAC library to isolate desired BAC clones, and we confirmed its utility using primer pairs designed for one of the clones. The Korean BAC library and screening pools will allow PCR-based screening of the Korean genome for any gene of interest. We also determined the allele types of HLA-DRA and HLA-DRB3 of clone KB55453, located in the HLA class II region on chromosome 6p21.3. The HLA-DRA and DRB3 genes in this clone were identified as the DRA*010202 and DRB3*01010201 types, respectively. The haplotype found in this library will provide useful information in future human disease studies.

Molecular Cloning and Characterization of a Novel Cold-Adapted Family VIII Esterase from a Biogas Slurry Metagenomic Library

  • Cheng, Xiaojie;Wang, Xuming;Qiu, Tianlei;Yuan, Mei;Sun, Jianguang;Gao, Junlian
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1484-1489
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    • 2014
  • A novel esterase gene, est01, was successfully unearthed from a biogas digester microbiota metagenomic library. The 1,194 bp est01 gene encodes a protein of 44,804 Da (designated Est01). The amino acid sequence of Est01 shows only moderate (33%) identity to a lipase/esterase. Phylogenetic analysis and biochemical characterization confirmed that Est01 is a new member of family VIII esterases. The purified Est01 from recombinant Escherichia coli BL21 (DE3) showed high hydrolytic activity against short-chain fatty acid esters, suggesting that it is a typical carboxylesterase rather than a lipase. Furthermore, the Est01 was even active at $10^{\circ}C$ (43% activity remained), with the optimal temperature at $20^{\circ}C$, and had a broad pH range from 5.0 to 10.0, with the optimal pH of 8.0. These properties suggest that Est01 is a cold-adaptive esterase and could have good potential for low-temperature hydrolysis application.

Phase Error Accumulation Methodology for On-chip Cell Characterization (온 칩 셀 특성을 위한 위상 오차 축적 기법)

  • Kang, Chang-Soo;Im, In-Ho
    • 전자공학회논문지 IE
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    • v.48 no.2
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    • pp.6-11
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    • 2011
  • This paper describes the design of new method of propagation delay measurement in micro and nanostructures during characterization of ASIC standard library cell. Providing more accuracy timing information about library cell (NOR, AND, XOR, etc.) to the design team we can improve a quality of timing analysis inside of ASIC design flow process. Also, this information could be very useful for semiconductor foundry team to make correction in technology process. By comparison of the propagation delay in the CMOS element and result of analog SPICE simulation, we can make assumptions about accuracy and quality of the transistor's parameters. Physical implementation of phase error accumulation method(PHEAM) can be easy integrated at the same chip as close as possible to the device under test(DUT). It was implemented as digital IP core for semiconductor manufacturing process($0.11{\mu}m$, GL130SB). Specialized method helps to observe the propagation time delay in one element of the standard-cell library with up-to picoseconds accuracy and less. Thus, the special useful solutions for VLSI schematic-to-parameters extraction (STPE), basic cell layout verification, design simulation and verification are announced.

Characterization of a Phage Library Displaying Random 22mer Peptides

  • Lee, Seung-Joo;Lee, Jeong-Hwan;Kay, Brian K.;Dreyfuss, Gideon;Park, Yong-Keun;Kim, Jeong-Kook
    • Journal of Microbiology
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    • v.35 no.4
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    • pp.347-353
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    • 1997
  • We have characterized a phage library displaying random 22mer peptides which were produced as N-terminal fusions to the pIII coat protein of M13 filamentous phages. Among the sixty phages randomly picked from the library, 25 phages had the 22mer peptide inserts. The DNA sequence analysis of the 25 inserts showed the following results: first, each nucleotide was represented almost equally at each codon position except that there were some biases toward G bases at the first position of the codons. Secondly, the expected 47 sense codons were represented. The deduced amino acid sequences of the 25 inserts were analyzed to examine its diversity. Glycine and glutamate were the two most overrepresented residues above the expected value, whereas cysteine and threonine residues were underrepresented. The range of dicersity in dipeptide sequences showed that the amino acid residues were randomly distributed along the peptide insert. Acidic, basic, polar, and nonpolar amino acid residues were represented to the extent expected at most positions of the peptide inserts. The predicted isoelectric points and hydropathy indices of the 25 peptides showed that a variety of the peptide were represented in the library. These results indicate that this phage display library could be useful in fiuding ligands for a broad spectrum of receptors by affinity screening.

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Cloning of Epidermis-specific cDNAS Encoding a Lipid Transfer Protein and an Aldehyde Decarbonylase from Senecio odorus

  • Pyee, Jaeho
    • Journal of Plant Biology
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    • v.39 no.3
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    • pp.189-195
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    • 1996
  • The major cuticular components have been shown to be synthesized in the epidermis. Therefore, cloning of epidermis-specific genes could yield information to be used to isolate and characterize the enzymes involved in the cuticle biosynthesis. A subtractive cDNA library was prepared from Senecio odorus in which epidermis-specific cDNAs were enriched. Differential screening of the library using epidermal and non-epidermal probes revealed two cDNAs. One of them designated epi425 was identified, based on the sequence homology, as a member of a new class in the LTP gene family and the other clone designated epi23 as a gene encoding an aldehyde decarbonylase. Northern blot analyses showed that epi425 and epi23 cDNAs hybridized with a transcript of about 600 and 2, 100 nucleotides, respectively, from the epidermis but not from the non-epidermal tissues. Further characterization of these clones will provide more information on the mechanism of the cuticle biosynthesis.

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High-throughput Preparation and Characterization of Powder and Thin-film Library for Electrode Materials

  • Fujimoto, Kenjiro;Onoda, Kazuhiro;Ito, Shigeru
    • Proceedings of the Korean Powder Metallurgy Institute Conference
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    • 2006.09a
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    • pp.254-255
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    • 2006
  • Powder library of pseudo four components Li-Ni-Co-Ti compounds were prepared for exploring the composition region with the single phase of the layer-type structure by using combinatorial high-throuput preparation system "M-ist Combi" based on electrostatic spray deposition method. The new layer-type compounds were found wider composition region than the previous report. This process is promising way to find multi component functional materials.

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Isolation and Characterization of New Family Genes of DNA Damage in Fission Yeast

  • Choi, In-Soon
    • Environmental Mutagens and Carcinogens
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    • v.19 no.1
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    • pp.28-33
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    • 1999
  • The SNF2 family includes proteins from a variety of species with roles I cellular processes such as transcriptional regulation, recombination and various types of DNA repair. Several proteins with unknown function are also included in this family. Here, we report the cloning and characterization of hrp 2+ gene (helicase related gene from S. pombe) which was isolated by PCR amplication using the conserved domain of SNF2 motifs within the ERCC6 gene which encodes a protein involved in DNA excision repair. The hrp2+ gene was isolated by screening with yeast S. pombe genomic library. The isolated cloned contained 6.5 kb insert DNA. Southern blot analysis confirmed that S. pombe chromosome contains the same DNA as hrp2+ gene and this gene exists as a single copy in S. pombe genome. The 4.7 kb transcript of mRNA was identified by Northern blot. To examined the transcriptional regulation of hrp2+ gene, DNA damaging agents were treated. These results indicated that the hrp2+ gene may not be directly involved in DNA replication, but may be involved in damage response pathway.

Functional Metagenome Mining of Soil for a Novel Gentamicin Resistance Gene

  • Im, Hyunjoo;Kim, Kyung Mo;Lee, Sang-Heon;Ryu, Choong-Min
    • Journal of Microbiology and Biotechnology
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    • v.26 no.3
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    • pp.521-529
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    • 2016
  • Extensive use of antibiotics over recent decades has led to bacterial resistance against antibiotics, including gentamicin, one of the most effective aminoglycosides. The emergence of resistance is problematic for hospitals, since gentamicin is an important broad-spectrum antibiotic for the control of bacterial pathogens in the clinic. Previous study to identify gentamicin resistance genes from environmental samples have been conducted using culture-dependent screening methods. To overcome these limitations, we employed a metagenome-based culture-independent protocol to identify gentamicin resistance genes. Through functional screening of metagenome libraries derived from soil samples, a fosmid clone was selected as it conferred strong gentamicin resistance. To identify a specific functioning gene conferring gentamicin resistance from a selected fosmid clone (35-40 kb), a shot-gun library was constructed and four shot-gun clones (2-3 kb) were selected. Further characterization of these clones revealed that they contained sequences similar to that of the RNA ligase, T4 rnlA that is known as a toxin gene. The overexpression of the rnlA-like gene in Escherichia coli increased gentamicin resistance, indicating that this toxin gene modulates this trait. The results of our metagenome library analysis suggest that the rnlA-like gene may represent a new class of gentamicin resistance genes in pathogenic bacteria. In addition, we demonstrate that the soil metagenome can provide an important resource for the identification of antibiotic resistance genes, which are valuable molecular targets in efforts to overcome antibiotic resistance.