• Title/Summary/Keyword: signal sequence

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Cloning of celC, Third Cellulase Gene, from Pectobacterium carotovorum subsp. carotovorum LY34 and its Comparison to Those of Pectobacterium sp.

  • LIM WOO JIN;RYU SUNG KEE;PARK SANG RYEOL;KIM MIN KEUN;AN CHANG LONG;HONG SU YOUNG;SHIN EUN CHULE;LEE JONG YEOUL;LIM YONG PYO;YUN HAN DAE
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.302-309
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    • 2005
  • Phytopathogenic Pectobacterium carotovorum subsp. carotovorum (Pcc) LY34 secretes multiple isozymes of the plant cell wall degrading enzyme endoglucanases. We have cloned a third cel gene encoding CMCase from Pcc LY34. The structural organization of the celC gene (AY188753) consisted of an open reading frame (ORP) of 1,116 bp encoding 371 amino acid residues with a signal peptide of 22 amino acids within the NH$_2$-terminal region of pre-CelC. The predicted amino acid sequence of CelC was similar to that of Peetobaeterium ehrysanthemi Cel8Y (AF282321). The CelC has the conserved region of the glycoside hydrolase family 8. The apparent molecular mass of CelC was calculated to be 39 kDa by CMC-SDS-PAGE. The cellulase­minus mutant of Pee LY34 was as virulent as the wild-type in pathogenicity tests on tubers of potato. The results suggest that the CelC of Pce LY34 is a minor factor for the pathogenesis of soft-rot.

Identification of Genetic Variations in CBL, SORBS1, CRK, and RHOQ, Key Modulators in the CAP/TC10 Pathway of Insulin Signal Transduction, and Their Association with Type 2 Diabetes Mellitus in the Korean Population

  • Hong, Kyung-Won;Jin, Hyun-Seok;Lim, Ji-Eun;Go, Min-Jin;Lee, Jong-Young;Hwang, Sue-Yun;Park, Hun-Kuk;Oh, Berm-Seok
    • Genomics & Informatics
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    • v.7 no.2
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    • pp.53-56
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    • 2009
  • Recent evidence has strongly suggested that the CAP/TC10 pathway is involved in the trafficking, docking, and fusion of vesicles containing the insulin-responsive glucose transporter Glut4 to the plasma membrane. However, little is known about how the genes employed in the CAP/TC10 pathway are associated with the development of type 2 diabetes mellitus. In this study, we sequenced 4 genes of the CAP/TC10 pathway [SORBS1, CBL, CRK, and RHOQ] in 24 individuals to identify genetic variations in these loci. A total of 48 sequence variants were identified, including 23 novel variations. To investigate the possible association with type 2 diabetes mellitus, 3 single nucleotide polymorphisms from SORBS1, 3 from CBL, and 4 from RHOQ were genotyped in 1122 Korean type 2 diabetic patients and 1138 nondiabetic controls. Using logistic regression analysis, 1 significant association between SNP rs1376405 in RHOQ and type 2 diabetes mellitus [OR = 8.714 (C.I. 1.714-44.29), p = 0.009] was found in the recessive model. Our data demonstrate a positive association of the RHOQ gene in the CAP/TC10 pathway with T2DM in the Korean population.

Performance evaluation of hybrid acquisition in CDMA systems (DS/CDMA 시스템에서 하이브리드 동기 획득의 성능 분석)

  • 강법주;강창언
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.23 no.4
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    • pp.914-925
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    • 1998
  • This paper considers the evaluation of the hybrid acquistion perdformance for the pilot signal in the direct sequence code division multiple access(DS/CDMA) forward link. the hybrid acquisition is introduced by the combination of two schemes, the parallel and serial acquisions. The mean acquisition time of the proposed scheme is derived to consider both the best case(the correct code-phase offsets are included i one subset) and the worst case(the correct code-phase offsets exist at the boundary of two subsets), which are cause by the distribution of the correct code-phase offsets in the subset. Expressions for the detection, false alarm, and miss probabilities are derived for the case of multiple correct code-phase offsets and multipath Rayleigh fading channel. Numerical results present the hybrid acquistion performance with repect to design parameters such as postdetectio integration length in the search and verification modes, subset size, and number of I/Q noncoherent correlators, and compare the hybrid acquistion with the parallel acquistion in terms of the minimum acquistion time under the same hardware complexity.

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Performance Evaluation of Hybrid DS/SFH-CDMA Noncoherent MFSK Signal with Channel Coding and MRC Diversity Techniques in Mobile Communication Nakagami Fading Channels (이동통신 Nakagami 페이딩 채널에서 채널코딩과 최대비합성 다이버시티 기법에 의한 Hybrid DS / SFH-CDMA 비동기 MFSK 신호의 성능평가)

  • ;Norihiko Morinaga
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.8 no.4
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    • pp.342-353
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    • 1997
  • This paper presents an analytical evaluation of a hybrid direct-sequence/slow frequencyhopped code division multiple-access (DS/SFH-CDMA) system employing noncoherent M-ary frequency shift keying(MFSK) modulation in a multiple Nakagami fading (m) environment. Multipath interference (MPI) and multi-access interference (MAI) is taken into account and the spectral efficiency is calculated for uncoded as well as channel coding systems. Predetection multipath maximal ratio combining (MRC) diversity in conjunction with interleaved channel coding (Hamming(7,4) code, BCH(15, 7) code and RS (7, 4), (15, 9)) code ) is employed for improving the bit error rate (BER) performance. The BER of noncoherent hybrid system is obtained using a Gaussian interference approximation.

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Performance Analysis of Diversity Received DS-CDMA/MDPSK Signals in m-distribution and Rician Fading Environments (m-분포 페이딩과 라이시안 페이딩 환경 하에서의 다이버시티 수신된 DS-CDMA / MDPSK 신호의 성능 분석)

  • 이정도;강희조
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.9 no.2
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    • pp.191-198
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    • 1998
  • The error are equation of DC-CDMA/MDPSK signal has been derived in m-distribution and Rician fading channels. Predeteception multipath MRC(Maximal Ratio Combining) diversity technique is employed for improving the bit error rate performance. The suitability of modeling a Rician fading environment by properly chosen m-distribution model is examined. Using the derived equation the error performance has been evaluated and shown in figures as a function of PN code sequence length(N), user number(U), multipath number(P), fading index(m), Rician factor(K), number of diversity branches(L) and ($E_b/N_o$). The results show that the error performance in Rician fading agrees well with that in m-distribution fading as fading becomes weak and as user number(U) increases and as multipath number(P) increases and diversity number(L) increases.

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Characteristics of Magnetic Resonance Imaging Findings in 32 Dogs Diagnosed with Meningoencephalitis of Unknown Etiology

  • Im, Chang-Gyu;Kim, Ah Reum;Han, Changhee;Hwang, Gunha;Kim, Rakhoon;An, Soyon;Hwang, Tae Sung;Lee, Hee Chun
    • Journal of Veterinary Clinics
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    • v.37 no.5
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    • pp.255-260
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    • 2020
  • The purpose of this retrospective study was to describe the characteristics of magnetic resonance imaging (MRI) findings in dogs with meningoencephalitis of unknown etiology (MUE), and to evaluate the usefulness of meningeal enhancement. Thirty-two dogs were included in MUE group on the basis of clinical signs, MRI findings and cerebrospinal fluid (CSF) results, and for comparison of the meningeal enhancement, twenty-three dogs with normal MRI, normal CSF and no clinical sign were included in the control group. The evaluated MRI findings included lesion site, lesion number, signal intensity of each MRI sequence, mass effect, perilesional edema, contrast enhancement, and meningeal enhancement. The MUE was most frequently associated with multiple lesions (50%) with perilesional edema (72%) in forebrain (66%) that were hyperintense (92%) in T2W and FLAIR images. Of the meningeal enhancement, there was no significant difference between the control group and the MUE groups in the pachymeningeal enhancement. However, leptomeningeal (or both) enhancement was found relatively high proportion in the MUE group than in the control group (P < 0.001, Odd ratio = 10.26), and based on this result, leptomeningeal (or both) enhancement is considered to be significant finding for indicating MUE.

Expression of Neurotensin/Neuromedin N Precursor in Murine Mast Cells

  • Ahn, Hyun-Jong;Cho, Jeong-Je
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.6
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    • pp.495-501
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    • 2001
  • We have cloned the mouse neurotensin/neuromedin N (NT/N) gene from the murine mast cell line Cl.MC/C57.1 for the first time. The murine NT/N cDNA clone consisted of 765 nucleotides and coded for 169 peptide residues with an N-terminal signal peptide, and the C-terminal region contained of one copy of neurotensin (NT) and one copy of neuromedin N (NN). Total of four Lys-Arg dibasic motifs were present; one each at the middle of the open reading frame, at the N-terminal of NN, at the C-terminal of NT, and between NN and NT. Amino acid sequence analysis of the mouse NT/N revealed 90% homology to that of the rat NT/N gene. NT/N is expressed in murine mast cell lines (Cl.MC/C57.1 and P815), but not in murine bone marrow-derived mast cells (BMMCs), murine macrophage cell line (RAW 264.7), nor in murine T cell line (EL-4). NT/N mRNA in C1.MC/C57.1 is highly inducible by IgE cross-linking, phorbol myristate acetate, neurotensin, and substance P. Following the treatment of demethylating agent, 5-azacytidine (5-azaC), the NT/N gene was induced in BMMCs in response to IgE cross-linking. 5-azaC-treated BMMCs did not express the NT/N gene without additional stimuli. These findings suggested that the regulation of NT/N gene expression was dependent on the effects of not only gene methylation but also enhancer and/or repressor proteins acting on the NT/N promoter.

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The Heavy Metal Tolerant Soil Bacterium Achromobacter sp. AO22 Contains a Unique Copper Homeostasis Locus and Two mer Operons

  • Ng, Shee Ping;Palombo, Enzo A.;Bhave, Mrinal
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.742-753
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    • 2012
  • Copper-containing compounds are introduced into the environment through agricultural chemicals, mining, and metal industries and cause severe detrimental effects on ecosystems. Certain microorganisms exposed to these stressors exhibit molecular mechanisms to maintain intracellular copper homeostasis and avoid toxicity. We have previously reported that the soil bacterial isolate Achromobacter sp. AO22 is multi-heavy metal tolerant and exhibits a mer operon associated with a Tn21 type transposon. The present study reports that AO22 also hosts a unique cop locus encoding copper homeostasis determinants. The putative cop genes were amplified from the strain AO22 using degenerate primers based on reported cop and pco sequences, and a constructed 10,552 base pair contig (GenBank Accession No. GU929214). BLAST analyses of the sequence revealed a unique cop locus of 10 complete open reading frames, designated copSRABGOFCDK, with unusual separation of copCD from copAB. The promoter areas exhibit two putative cop boxes, and copRS appear to be transcribed divergently from other genes. The putative protein CopA may be a copper oxidase involved in export to the periplasm, CopB is likely extracytoplasmic, CopC may be periplasmic, CopD is cytoplasmic/inner membrane, CopF is a P-type ATPase, and CopG, CopO, and CopK are likely copper chaperones. CopA, B, C, and D exhibit several potential copper ligands and CopS and CopR exhibit features of two-component regulatory systems. Sequences flanking indicate the AO22 cop locus may be present within a genomic island. Achromobacter sp. strain AO22 is thus an ideal candidate for understanding copper homeostasis mechanisms and exploiting them for copper biosensor or biosorption systems.

Cloning and Expression of a Thermostable ${\alpha}$-Galactosidase from the Thermophilic Fungus Talaromyces emersonii in the Methylotrophic Yeast Pichia pastoris

  • Simila, Janika;Gernig, Anita;Murray, Patrick;Fernandes, Sara;Tuohy, Maria G.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1653-1663
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    • 2010
  • The first gene (${\alpha}$-gal1) encoding an extracellular ${\alpha}$-Dgalactosidase from the thermophilic fungus Talaromyces emersonii was cloned and characterized. The ${\alpha}$-gal1 gene consisted of an open reading frame of 1,792 base pairs interrupted by six introns that encoded a mature protein of 452 amino acids, including a 24 amino acid secretory signal sequence. The translated protein had highest identity with other fungal ${\alpha}$-galactosidases belonging to glycosyl hydrolase family 27. The ${\alpha}$-gal1 gene was overexpressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast Pichia pastoris. Recombinant ${\alpha}$-Gal1 was secreted into the culture medium as a monomeric glycoprotein with a maximal yield of 10.75 mg/l and purified to homogeneity using Hisbinding nickel-agarose affinity chromatography. The purified enzyme was maximally active at $70^{\circ}C$, pH 4.5, and lost no activity over 10 days at $50^{\circ}C$. ${\alpha}$-Gal1 followed Michaelis-Menten kinetics ($V_{max}\;of\;240.3{\mu}M/min/mg,\;K_m\;of\;0.294 mM$) and was inhibited competitively by galactose ($K_m{^{obs}}$ of 0.57 mM, $K_i$ of 2.77 mM). The recombinant T. emersonii ${\alpha}$-galactosidase displayed broad substrate preference, being active on both oligo- and polymeric substrates, yet had strict specificity for the ${\alpha}$-galactosidic linkage. Owing to its substrate preference and noteworthy stability, ${\alpha}$-Gal1 is of particular interest for possible biotechnological applications involving the processing of plant materials.

Construction of a Shuttle Vector for Protein Secretory Expression in Bacillus subtilis and the Application of the Mannanase Functional Heterologous Expression

  • Guo, Su;Tang, Jia-Jie;Wei, Dong-Zhi;Wei, Wei
    • Journal of Microbiology and Biotechnology
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    • v.24 no.4
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    • pp.431-439
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    • 2014
  • We report the construction of two Bacillus subtilis expression vectors, pBNS1/pBNS2. Both vectors are based on the strong promoter P43 and the ampicillin resistance gene expression cassette. Additionally, a fragment with the Shine-Dalgarno sequence and a multiple cloning site (BamHI, SalI, SacI, XhoI, PstI, SphI) were inserted. The coding region for the amyQ (encoding an amylase) signal peptide was fused to the promoter P43 of pBNS1 to construct the secreted expression vector pBNS2. The applicability of vectors was tested by first generating the expression vectors pBNS1-GFP/pBNS2-GFP and then detecting for green fluorescent protein gene expression. Next, the mannanase gene from B. pumilus Nsic-2 was fused to vector pBNS2 and we measured the mannanase activity in the supernatant. The mannanase total enzyme activity was 8.65 U/ml, which was 6 times higher than that of the parent strain. Our work provides a feasible way to achieve an effective transformation system for gene expression in B. subtilis and is the first report to achieve B. pumilus mannanase secretory expression in B. subtilis.