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A Novel Cellulase of the Mulberry Longicorn Beetle, Apriona germari, Dependent on N-Glycosylation for Enzymatic Activity

  • Lee, Seong-Jin;Kim, Seong-Ryul;Yoon, Hyung-Joo;Kim, IK-Soo;Lee, Kwang-Sik;Je, Yeon-Ho;Lee, Sang-Mong;Seo, Sook-Jae;Sohn, Hung-Dae;Jin, Byung-Rae
    • 한국잠사학회:학술대회논문집
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    • 한국잠사학회 2003년도 제46회 춘계 학술연구 발표회
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    • pp.77-78
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    • 2003
  • A novel -1, 4-endoglucanase (EGase, EC 3.2.1.4) cDNA belonging to glycoside hydrolase family (GHF) 45 was cloned from the mulberry longicorn beetle, Apriona germari. The cDNA encoding EGase of A. germari (Ag-EGase) is 711 base pairs long with an open reading frame of 237 amino acid residues. The deduced protein sequence of Ag-EGase showed 54% and 48% identity to phytophagous beetle Phaedon cochleariae and termite Reticulitermes speratus hindgut symbiont, respectively. (omitted)

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Characterization of the Small Cryptic Plasmid, pGD2, of Klebsiellia sp. KCL-2.

  • Yoo, Ju-Soon;Kim, Hae-Sun;Chung, Soo-Yeol;Lee, Young-Choon;Cho, Young-Soo;Choi, Yong-Lark
    • BMB Reports
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    • 제34권6호
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    • pp.584-589
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    • 2001
  • One of the cryptic plasmids from the oil degrading bacterium Klebsiella sp. KCL-2, the small plasmid pGD2, has been identified and characterized. This plasmid has a size of 3.6 kb with unknown functions. We constructed the recombinant plasmid pMGD2. The nucleotide sequences of the plasmid were determined and two open reading frames were detected. ORF1 encodes a replication initiator protein (RepA), which has a high degree of homology with the protein of ColE2 plasmid. The product encoded by ORF2 showed a high similarity with the transposase protein of IS5. IS5 is 1195 by long and contains an inverted terminal repetition of 16 bp with one mismatch. Stem-loop structures in the 5'untranslated region of the repA suggest that a putative gene, incA, is located in a complementary strand to the leader region of the repA mRNA.

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Molecular Cloning and Characterization of Very Late Expression Factor 1 Gene, vlf-1 from Bombyx mori Nuclear Polyhedrosis Virus Kl

  • Park, Hye-Jin;Lee, Kwang-Sik;Cho, Eun-Sook;Yun, Eun-Young;Kang, Seok-Woo;Kim, Keun-Young;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제1권1호
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    • pp.29-33
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    • 2000
  • We cloned and characterized a very late expression factor 1 gene, vlf-1, which regulates the level of very late gene transcripts, from Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain. The 1,140 bp vlf-1 has an open reading frame of 379 amino acid and a MW of 44 kDa. The vlf-1 nucleotide sequence of BmNPV-Kl showed high homology with Autographa californica nuclear polyhedrosis virus and BmNPV T3 strain so far known, and its deduced amino acid residues were identical to those of BmNPV T3. The location of vlf-1 in the BmNPV-Kl genome was confirmed by Southern blot analysis and its expression patterns at the transcriptional level were confirmed by Northern hybridization analysis.

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cDNA Sequence of a Novel Immulectin Homologue from the Silkworm, Bombyx mori

  • Kim, Seong-Ryul;Lee, Kwang-Sik;Kim, Iksoo;Kang, Seok-Woo;Nho, Si-Kab;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제6권1호
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    • pp.99-102
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    • 2003
  • A cDNA of novel immulectin homologue (BmIML), a C-type lectin, was cloned from the silkworm, Bombyx mori. The immulectin cDNA is an open reading frame of 921 bp encoding 307 amino acid residues. The deduced amino acid sequence from the BmIML cDNA contains two C-type carbohydrate recognition domains (CRDs). The BmIML was most similar (61 % protein sequence identity) to the M. sexta immulectin-1, whereas BmIML showed relatively lower identity to the B. mori lipopolysaccharide-binding protein (25% protein sequence identity). These features of BmIML indicate that BmIML is a novel member of C-type lectin superfamily. Northern blot analysis revealed that the BmIML is specifically expressed in the fat body of B. moli larvae.

Molecular Cloning of a LIM Protein cDNA from the Mulberry Longicorn Beetle, Apriona germari

  • Gui, Zhongzheng;Wei, Yadong;Yoon, Hyung Joo;Kim, Iksoo;Guo, Xijie;Jin, Byung Rae;Sohn, Hung Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제9권1호
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    • pp.149-153
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    • 2004
  • Here we report the molecular cloning of a LIM protein cDNA of the CRP (cysteine-rich protein) family from the mulberry longicorn beetle, Apriona, geramri. The A. germari LIM protein cDNA contains an open reading frame of 276 bp encoding 92 amino acid residues with a calculated molecular weight of approximately 10 kDa. The A. germari LIM protein contains the cysteine-rich consensus sequence of LIM domain and the glycine-rich consensus sequence observed in cysteine-rich protein family 1 (CRP1). The potential nuclear targeting signal is retained. The deduced amino acid sequence of the A. germari LIM protein cDNA showed 81 % identity to both Bombyx mori muscle LIM protein (Mlp) and Drosophila melanogaster Mlp60A and 77% to Epiblema scudderiana Mlp. Northern blot analysis showed that A. germari LIM protein is highly expressed in epidermis and muscle, and less strongly in midgut, but not in the fat body.

Molecular Cloning of a cDNA Encoding a Cathepsin B Homologue from the Mulberry Longicorn Beetle, Apriona germari

  • Kim, Seong-Ryul;Yoon, Hyung-Joo;Park, Nam-Sook;Lee, Sang-Mong;Moon, Jae-Yu;Jin, Byung-Rae;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제4권1호
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    • pp.63-68
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    • 2002
  • A cDNA encoding a putative member of cathepsin B of the thiol pretense superfamily was cloned from a cDNA library of the mulberry longicorn beetle, Apriona germari. Sequence analysis of the cDNA encoding the cathepsin B of A. germari (AgCatB) revealed that the 972 bp cDNA has an open reading frame of 324 amino acid residues. The deduced protein sequence of the AgCatB showed high homology with cathepsin B of the insects, Bombyx mori (47.3% amino acid identity), Helicoverpa armigera (46.6%) and Sarcophaga peregrina (45.6%), and the lowest homology with Aedes aegypti (33.2%). The AgCatB contains six disulfate bonds typical for cysteine pretenses. The three amino acid positions Cys-109, His-267, and Asn-287 which are conserved, active sites characteristic for cathepsin B, were also found. Phylogenetic analysis further confirmed that the AgCatB has a close relationship with that of B. mori, H. armigera and S. peregrina.

Molecular Characterization of the Genes Encoding Acetoacetyl-Coenzyme A Transferase from Serratia marcescens KCTC 2172

  • Yoo, Ju-Soon;Kim, Hae-Sun;Lee, Young-Choon;Chung, Soo-Yeol;Choi, Yong-Lark
    • Journal of Microbiology and Biotechnology
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    • 제11권5호
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    • pp.870-875
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    • 2001
  • A DNA fragment, pCKB13, containing two genes encoding Coenzyme a transferase, was isolated from a genomic DNA library of S. marcescens KCTC 2172. The complete nucleotide sequence of the 2,081-bp BamHI fragment on pCKB13 was determined. Sequencing of the fragment led to the identification of two open reading frames showing high homology with two Coenzyme A (CoA) transferases, Acetoacetyl-CoA transferase (Acot) and Succinyl-CoA transferase (Scot), enzymes catalyzing the reversible transfer of CoA from one carboxylic acid to another. The enzyme activity of Coenzyme A transferase increased after introducing the multicopy of the cloned gene in E. coli. The recombinant protein, overexpressed by multicopy and induction with IPTG, was a polypeptide of 42 kDa, as confirmed by SDS-PAGE. The protein was purified to homogeneity through three sequential chromatographic procedures including ion-exchanged DEAE-sepharose, CM-sepharose, and Mono Q.

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Isolation and Characterization of cDNA Encoding Pyridoxal Kinase from Ovine Liver

  • Lee, Hyun-Shik;Choi, Soo-Young;Kwon, Oh-Shin
    • BMB Reports
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    • 제32권5호
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    • pp.502-505
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    • 1999
  • cDNA fragments of ovine liver pyridoxal kinase were amplified by PCR using degenerate oligonucleotide primers derived from partial amino acids sequences of the enzyme. Using PCR products as probes, several overlapping cDNA clones were isolated independently from an ovine liver and a human brain cDNA library. The largest cDNA clone for each was selected for sequence analysis. The ovine liver cDNA encodes a polypeptide of 297 amino acid residues with Mr of 32,925, whereas the human clone is comprised of an open reading frame encoding 312 amino acid residues with Mr of 35,102. The deduced sequence of the human brain enzyme is completely identical to that of human testes cDNA recently reported (Hanna et al., 1997). The ovine enzymes have approximately 77% sequence identity with the human enzyme although the two sequences are completely different in the N-terminus comprising 32 residues. This result suggests that pyridoxal kinase is highly homologous in mammalian species.

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Molecular Cloning and Sequencing of Cell Wall Hydrolase Gene of an Alkalophilic Bacillus subtilis BL-29

  • Kim, Tae-Ho;Hong, Soon-Duck
    • Journal of Microbiology and Biotechnology
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    • 제7권4호
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    • pp.223-228
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    • 1997
  • A DNA fragment containing the gene for cell wall hydrolase of alkalophilic Bacillus subtilis BL-29 was cloned into E. coli JM109 using pUC18 as a vector. A recombinant plasmid, designated pCWL45B, was contained in the fragment originating from the alkalophilic B. subtilis BL-29 chromosomal DNA by Southern hybridization analysis. The nucleotide sequence of a 1.6-kb HindIII fragment containing a cell wall hydrolase-encoding gene was determined. The nucleotide sequence revealed an open reading frame (ORF) of 900 bp with a concensus ribosome-binding site located 6 nucleotide upstream from the ATG start codon. The primary amino acid sequence deduced from the nucleotide sequence revealed a putative protein of 299 amino acid residues with an M.W. of 33, 206. Based on comparison of the amino acid sequence of the ORF with amino acid sequences in the GenBank data, it showed significant homology to the sequence of cell wall amidase of the PBSX bacteriophage of B. subtilis.

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일정체적 변단면 기둥의 자유진동 (Free Vibrations of Tapered Columns with Constant Volume)

  • 이병구;이태은;최규문;송주한
    • 한국소음진동공학회:학술대회논문집
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    • 한국소음진동공학회 2002년도 춘계학술대회논문집
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    • pp.417-422
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    • 2002
  • The main purpose of this paper is to determine the dynamic optimal shapes of tapered column with constant volume. The linear, parabolic and sinusoidal tapers with the regular polygon cross-section are considered, whose material volume and span length are always held constant. The ordinary differential equation including the effect of axial load is applied to calculate the natural frequencies. The Runge-Kutta method and Regula-Falsi methods are used to integrate the differential equation and compute the frequencies, respectively. Then the dynamic optimal shape whose lowest natural frequency is highest is determined by reading the critical value of the frequency versus section ratio curve plotted by the frequency data. In the numerical examples, the tapered columns are analysed and the numerical result of this study are shown in table and figures.

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