• Title/Summary/Keyword: Gene bank

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Expression of Thermostable $\alpha$-Glucosidase from Thermus caldophilus GK24 in Recombinant Saccharomyces cerevisiae

  • Choi, Jae-Youl;Ahn, Jung-Oh;Kim, Sun-Il;Shin, Hyun-Jae
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.2000-2003
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    • 2006
  • A gene (GenBank AF096282) coding for a $\alpha$-glucosidase (TcaAG, EC 3.2.1.20) from Thermus caldophilus GK24 was expressed in Saccharomyces cerevisiae, a generally recognized as safe (GRAS) host. The thermostable $\alpha$-glucosidase was produced inside of the GRAS host at 0.04 unit/mg-dry cell by the constitutively expressing ADH1 promoter and at 1.2 unit/mg-dry cell by the inductively expressing GALl0 promoter, respectively. No $\alpha$-glucosidase activities were found in the medium when the MF-alpha signal sequence from S. cerevisiae or $\alpha$-amylase signal sequence from Aspergillus oryzae were fused before the $\alpha$-glucosidase gene for the secretion.

Development of Information Biology (I)

  • Tateno, Yoshio
    • Interdisciplinary Bio Central
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    • v.5 no.1
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    • pp.2.1-2.3
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    • 2013
  • Birth and development of information biology are introduced with its definition and scientific basis. The discipline lives on the two types of nutrition, one is a huge amount of biological data on genomes, gene expressions, proteomes, protein 3D structures, protein networks, and so forth. The other is the method of using them on a computer. The scientific basis of the two is evolution. To collect genome and gene expression data form laboratories in the world, annotate and dissimilate back to researchers worldwide, they built the EMBL database in Europe in 1982, GenBank in USA in 1984 and DNA Data Bank of Japan in 1987. On the other hand, the methods of using and analyzing those data have accordingly been developed. The two aspects advance the discipline further and further.

Eriodictyol induces apoptosis via regulating phosphorylation of JNK, ERK, and FAK/AKT in pancreatic cancer cells

  • Oh, Ui Hyeon;Kim, Da-Hye;Lee, Jungwhoi;Han, Song-I;Kim, Jae-Hoon
    • Journal of Applied Biological Chemistry
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    • v.65 no.2
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    • pp.83-88
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    • 2022
  • Although it has been intensively studied over the past few decades, pancreatic cancer remains one of the most lethal cancers. Eriodictyol, a plant-derived flavonoid mainly found in citrus fruits, exerts diverse biological effects, including anti-oxidant, anti-cancer, and anti-inflammatory properties. In this study, we investigated the anticancer properties of eriodictyol and its mechanisms of action in pancreatic cancer cells. In both SNU213 and Panc-1 cells, eriodictyol decreased viability, induced apoptosis, and decreased clonogenicity. In addition, eriodictyol treatment increased the phosphorylation level of JNK and decreased the phosphorylation levels of ERK, FAK, and AKT. These observations provide insight into the molecular mechanisms of eriodictyol-induced apoptosis in pancreatic cancer cell lines, and could contribute to the development of candidate compounds for treating pancreatic cancer.

Identification of Bak-like Protein cDNA (Bak-like 단백질을 code하는 cDNA의 동정)

  • 김진경
    • YAKHAK HOEJI
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    • v.45 no.4
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    • pp.426-430
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    • 2001
  • Cells are eliminated in a variety of physiological settings by apoptosis, a genetically encoded process of cellular suicide. Bak, a member of the Bcl-2 protein family, accelerates apoptosis by an unknown mechanism. We have found a novel cDNA encoding a 101 amino acid protein possessing a Bak-like in our full-length cDNA bank. Bak-like shares the conserved domains BHI and 2 with other proapoptotic proteins but lacks the BH3 domain. Bak-like is expressed in a wide variety of tissues. Like Bak, Bak-like gene product primarily enhances apoptotic cell death following an appropriate stimulus.

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Subspecific Status of the Korean Tiger Inferred by Ancient DNA Analysis

  • Lee, Mu-Yeong;Hyun, Jee-Yun;Lee, Seo-Jin;An, Jung-Hwa;Lee, Eun-Ok;Min, Mi-Sook;Kimura, Junpei;Kawada, Shin-Ichiro;Kurihara, Nozomi;Luo, Shu-Jin;O'Brien, Stephen J.;Johnson, Warren E.;Lee, Hang
    • Animal Systematics, Evolution and Diversity
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    • v.28 no.1
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    • pp.48-53
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    • 2012
  • The tiger population that once inhabited the Korean peninsula was initially considered a unique subspecies (Panthera tigris coreensis), distinct from the Amur tiger of the Russian Far East (P. t. altaica). However, in the following decades, the population of P. t. coreensis was classified as P. t. altaica and hence forth the two populations have been considered the same subspecies. From an ecological point of view, the classification of the Korean tiger population as P. t. altaica is a plausible conclusion. Historically, there were no major dispersal barriers between the Korean peninsula and the habitat of Amur tigers in Far Eastern Russia and northeastern China that might prevent gene flow, especially for a large carnivore with long-distance dispersal abilities. However, there has yet to be a genetic study to confirm the subspecific status of the Korean tiger. Bone samples from four tigers originally caught in the Korean peninsula were collected from two museums in Japan and the United States. Eight mitochondrial gene fragments were sequenced and compared to previously published tiger subspecies' mtDNA sequences to assess the phylogenetic relationship of the Korean tiger. Three individuals shared an identical haplotype with the Amur tigers. One specimen grouped with Malayan tigers, perhaps due to misidentification or mislabeling of the sample. Our results support the conclusion that the Korean tiger should be classified as P. t. altaica, which has important implications for the conservation and reintroduction of Korean tigers.

Functional assessment of attenuated mutants of Pepper mild mottle virus

  • Yoon, J.Y.;Tsuda, S.;Ryu, K.H.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.144.1-144
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    • 2003
  • Attenuated viruses can protect their hosts against challenge to their related viruses. Increasing evidence shows that mutations of the tobamoviral 126/183 kDa protein play a major role in the viral attenuation and contribute to the cross protection mechanism. In this study, four mutants of Pepper mild mottle virus (PMMoV) have been constructed by mutagenesis; two mutants, pTPpoly348 and pTPpoly762, were substituted in the middle of replicase gene, and the others, pTPL3D:: $\Delta$6207 and pTPL3D:: $\Delta$6219, were deletion mutants made by deleting some parts of pseudoknot structures of the 3' noncoding region (NCR) of the virus. Progeny viruses generated from the four mutants were infectious on N. benthamiana plants with symptomless or mild mosaic symptom. Replication efficiency and viral product accumulations of four mutants were assessed by Northern and Western blot analyses on BY-2 protoplast cells. Accumulation of CP for the pTPL3D:: $\Delta$6207 and pTPL3D:: $\Delta$6219 were lower than that of other mutants and wild type virus. These data suggest that the 3'-NCR mutations contribute to the viral gene expression in host tissues, while mutants of replicase gene rather govern the symptom expression.

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Cloning and Sequencing of a Novel Glutaryl Acylase ${\beta}-Subunit$ Gene of Pseudomonas cepacia BY21 from Bioinformatics

  • Jeong, Yoo-Seok;Yoo, Hyo-Jin;Kim, Sang-Dal;Nam, Doo-Hyun;Khang, Yong-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.510-515
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    • 2005
  • Pseudomonas cepacia BY21 was found to produce glutaryl acylase that is capable of deacylating glutaryl-7-aminocephalosporanic acid (glutaryl-7-ACA) to 7-aminocephalosporanic acid (7-ACA), which is a starting material for semi-synthetic cephalosporin antibiotics. Amino acids of the reported glutaryl acylases from various Pseudomonas sp. strains show a high similarity (>93% identity). Thus, with the known nucleotide sequences of Pseudomonas glutaryl acylases in GenBank, PCR primers were designed to clone a glutaryl acylase gene from P. cepacia BY21. The unknown -subunit gene of glutaryl acylase from chromosomal DNA of P. cepacia BY21 was cloned successfully by PCR. The -subunit amino acids of P. cepacia BY21 acylase (GenBank accession number AY948547) were similar to those of Pseudomonas diminuta KAC-1 acylase except that Asn408 of P. diuminuta KAC-1 acylase was changed to Leu408.

Molecular Characterization of Nippostrongylus brasiliensis (Nematoda: Heligmosomatidae) from Mus musculus in India

  • Chaudhary, Anshu;Goswami, Urvashi;Singh, Hridaya Shanker
    • Parasites, Hosts and Diseases
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    • v.54 no.6
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    • pp.743-750
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    • 2016
  • Mus musculus (Rodentia: Muridae) has generally been infected with a rodent hookworm Nippostrongylus brasiliensis. In this report, we present morphological and molecular identification of N. brasiliensis by light and scanning electron microscopy and PCR amplification of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene and the protein sequences encoded by cox1 gene, respectively. Despite the use of N. brasiliensis in many biochemistry studies from India, their taxonomic identification was not fully understood, especially at the species level, and no molecular data is available in GenBank from India. Sequence analysis of cox1 gene in this study revealed that the present specimen showed close identity with the same species available in GenBank, confirming that the species is N. brasiliensis. This study represents the first record of molecular identification of N. brasiliensis from India and the protein structure to better understand the comparative phylogenetic characteristics.

Isolation and Characterization of a Novel Triolein Selective Lipase from Soil Environmental Genes

  • Lim, Hee Kyung;Han, Ye-Jin;Hahm, Moon-Sun;Park, Soo Youl;Hwang, In Taek
    • Microbiology and Biotechnology Letters
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    • v.48 no.4
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    • pp.480-490
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    • 2020
  • A novel lipase gene, Lip-1420, was isolated from a metagenomic library constructed from reed marsh from Mt. Jumbong in Korea, comprising 112,500 members of recombinant plasmids. The DNA sequence of Lip-1420-subclone (5,513 bp) was found to contain at least 11 ORFs according to the GenBank database. The ORF-3 gene was inserted into the pET21a plasmid containing the C-terminal 6-His tag and transformed into E. coli BL21(DE3) to express the recombinant lipase protein. Lip-1420 was purified using a fast protein liquid chromatography system. The gene was registered in GenBank (MH628529). The values of Km and Vmax were determined as 0.268 mM and 1.821 units, respectively, at 40℃ and pH 8.0, using p-nitrophenyl palmitate as the substrate. This lipase belongs to family IV taxonomically because it has conserved HGGG and GDSAG motifs in the constitutive amino acid sequence. According to the predicted structural model, the binding sites are represented by residues H78, G81, D150, S151, A152, V181, and D236. Finally, Lip-1420 showed triolein selectivity for methanolysis between triolein (18:1) and tristearin (18:0) substrates. Further study of the selective mechanism and structure-function relationship of this new lipase could be useful for more practical applications.