• Biotechnology and Bioprocess Engineering:BBE
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• 제7권1호
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• pp.38-42
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• 2002

A chitinase gene (pCHi58) encoding a 58 kDa chitinase was isolated from the Serratia marcescens KCTC 2172 cosmid library. The chitinase gene consisted of a 1686 bp open reading frame that encoded 562 amino acids. Escherichia coil harboring the pChi58 gene secreted a 58 kDa chitinase into the culture supernatant. The 58 kDa chitinase was purified using a chitin affinity column and mono-S column. A nucleotide and N-terminal amino acid sequence analysis showed that the 58 kDa chitinase had a leader peptide consisting of 23 amino acids which was cleaved prior to the 24th alanine. The 58 KDa chitinase exhibited a $98\%$ similarity to that of S. marcescens OMB 1466 in its nuclotide sequence. The chitinolytic patterns of the 58 kDa chitinase released N,N'-diacetyl chitobiose (NAG2) as the major hydrolysis end-product with a trace amount of N-acetylglucosamine. When a 4-methylumbellyferyl-N-acetylglucosamin monomer, dimmer, and tetramer were used as substrates, the 58 kDa chitinase did not digest the 4-Mu-NAG monomer $(analogue\;of\;NAG_2)$, thereby indicating that the 58 kDa chitinase was likely an endochitinase. The optimum reaction temperature and pH of the enzyme were $50^{\circ}C$ and 5.0, respectively.

• 한국가금학회지
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• 제40권3호
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• pp.243-252
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• 2013

Among the 16 hemagglutinin (HA) subtypes of avian influenza virus (AIV), only the H5 and H7 subtypes have caused highly pathogenic avian influenza (HPAI) in poultry. However, most H5 or H7 subtype viruses are categorized as low pathogenic avian influenza (LPAI). Some AIVs, including the H5 and H7 HPAI viruses, have shown the ability to infect humans directly. In this study, we describe the biological and molecular characterization of an H5N3 AIV (SBD/KR/KNU SYG06/06) isolated from spot-billed duck (Anas poecilorhyncha) in Korea. A phylogenetic analysis of the eight viral genes showed that the SBD/KR/KNU SYG06/06 isolate belongs to the Eurasian lineage and that the SBD/KR/KNU SYG06/06 isolate was clearly different from HPAI H5N1 strains, including human isolates and the Italian HPAI H5N2 strains. Additionally, no relationship was found between SBD/KR/KNU SYG06/06 and the Korean HPAI H5N1 isolates. The SBD/KR/ KNU SYG06/06 isolate had avian specific receptor binding site residues in the HA protein and the four C-terminal amino acids in the NS1 protein. The HA protein of the SBD/KR/KNU SYG06/06 isolate exhibited the typical LPAI motif at the cleavage site and this virus produced no cytopathic effects in MDCK cells without trypsin. Given these results, we suggest that the H5N3 AIV isolated from the spot-billed duck should be considered an LPAI virus and should have no pathogenic effect in humans.

• 한국약용작물학회지
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• 제16권3호
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• pp.195-198
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• 2008

Karyotypic analysis and FISH (fluorescence in situ hybridization) with 45S and 5S rRNA genes were carried out in Persicaria tinctoria H Gross. The somatic metaphase chromosomes were ranged from 2.25 ${\mu}m$ to 1.50 ${\mu}m$ in length. Chromosome number was 2n = 4x = 40 with the basic number of x = 10. The chromosome complement of the species consisted of 16 pairs of metacentrics (chromososomes 1,2,3,4,6,7,8,9, 10, 11, 12, 13, 15, 18, 19 and 20) and 4 pairs of submetacentrics (chromosome 5, 14, 16 and 17). The karyotype formula was K(2n) = 4x = 32 m + 8 sm. In FISH analysis, three pairs of 45S rRNA gene loci on the terminal region of submetacentrics (chromosomes 5, 16 and 17) and two pairs of 5S rRNA gene loci on the centromeric region of metacentrics (chromosomes 9 and 11) were detected, respectively.

• Journal of Microbiology and Biotechnology
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• 제9권6호
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• pp.839-846
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• 1999

An extracellular chitinase-producing bacterial strain induced by colloidal chitin was isolated from sea sand and was identified to be a member of the genus Cytophaga. The chitinase was purified successively by 30-60% ammonium sulfate fractionation, and DEAE-Bio gel A column, Octyl-Sepharose CL-4B column, and DEAE-Bio gel A column chromatographies. The enzyme had a molecular mass of 59.75 kDa, and the amino terminal amino acid sequence was ATPNAPVISW MPTDXXLQNXS. The enzyme acted better on colloidal chitin as a substrate than on chitosan. For colloidal chitin and chitosan (Degree of Acetylation, 15-25%), $K_{cat}$ values were 0.60U/mg and 0.08U/mg, respectively. HPLC analysis of the enzymatic reaction products showed that the chitinase produced mostly N-acetyl-D-glucosarnine and di-N-acetylchitobiose. The optimum temperature and pH for the enzyme were $50^{\circ}C$ and 4.0, respectively. N-Bromosuccinimide and $Hg^{2+}$ inhibited the chitinase activity as much as 90%, and $Sb^{3+}$, diethylpyrocarbonate, and $Ag^{+}$ inhibited it by 50-70%.

• Applied Biological Chemistry
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• 제34권4호
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• pp.334-338
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• 1991

고상법으로 새로운 반응기구에 의한 bradykinin 및 $(D-Phe7\;-Leu^8)$ bradykinin을 합성하였다. Coupling은 N, N'-dicyclohexylcarbodiimide로 행하였으며 HBr 용액으로 cleavage한 후 조펩티드는 high pressure liquid chromatography로 정제하였다. 이들 펩티드의 순도는 paper chromatography, thin layer chromatography, paper electrophoresis, 융점측정기 및 아미노산기분석기에 의하여 분석하였다. Endopeptidase인 ${\alpha}-chymotrypsin$과 trysin, exopeptidase인 carboxypeptidase A와 leucine aminopeptidase를 사용하여 in vitro 상에서 이들 펩티드의 분해실험을 하였다. ${\alpha}-Chymotrypsine$ 및 carboxypeptidase A에 의하여 이들 펩티드는 빠르게 분해하였으나 leucine aminopeptidase는 N-말단의 2번 위치에 proline의 imino결합 때문에 분해하지 않았다.

• Molecules and Cells
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• 제20권1호
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• pp.57-68
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• 2005

Murine erythroleukemia (MEL) cells are widely used to study erythroid differentiation thanks to their ability to terminally differentiate in vitro in response to chemical induction. At the molecular level, not much is known of their terminal differentiation apart from activation of adult-type globin gene expression. We examined changes in gene expression during the terminal differentiation of these cells using microarray-based technology. We identified 180 genes whose expression changed significantly during differentiation. The microarray data were analyzed by hierarchical and k-means clustering and confirmed by semi-quantitative RT-PCR. We identified several genes including H1f0, Bnip3, Mgl2, ST7L, and Cbll1 that could be useful markers for erythropoiesis. These genetic markers should be a valuable resource both as potential regulators in functional studies of erythroid differentiation, and as straightforward cell type markers.

• Molecules and Cells
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• 제24권1호
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• pp.148-151
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• 2007

Porcine endogenous retroviruses (PERVs) in the pig genome represent a potential risk of infection in pig-to-human transplantation and are transmitted vertically. The solitary long terminal repeat (LTR) elements of the PERVs affect the replication properties of the individual viruses via their repeat sequences and by encoding a set of specific transcription factors. We examined the promoter activities of solitary LTR elements belonging to the PERV-A and -B families of the Korean domestic pig (KDP) using luciferase reporters. Three of the LTR structures (of PERV-A5-KDP, PERV-A7-KDP, PERV-A8-KDP) had different promoter activities in human HCT116 cells and monkey Cos7 cells, and potential negatively and positively acting regions affecting transcription were identified by deletion analysis. These data suggest that specific sequences in the U3 region of a given LTR element can affect the activities of promoter or enhancer elements in the PERV.

• 디지털산업정보학회논문지
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• 제10권3호
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• pp.11-21
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• 2014

In this paper, concerning the various advertising and policy advertising of the election with respect to whether to deliver a message to a large number of people, we design and implement an automative system what enables sending the text messages directly from the server to the client and also fast feedback is enabled by utilizing a number of operational programs to connect to the server. Therefore, we design and implement the automative communication system which enables delivering message to each user mobile terminal from a plurality of relay mobile terminals by utilizing the mobile instant messenger, not to deliver a message from the server to the mobile instant messenger user directly. In result of comparative analysis on the number of times of data transmission, this automative communication system utilizing mobile instant messenger shows the result that it enables transmitting five times per minute as it can copy and paste in the automation system regardless of the size of the data loading, otherwise in case of transmitting manually it show the result that the number of times of data transmission is reduced if the size of the data is larger.

• 미생물학회지
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• 제35권1호
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• pp.18-24
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• 1999

Kinesin은 Saccharomyces cerevisiae, Aspergillus nidulans, Drosophila melanogaster 등에서 발견 되었으며 dynein 과 더불어 세포분열 과정중 chromosome 의 이동과 spindle pole 의 분리에 관련된 것으로 알려져 있다. 본 연구에서는 기존에 발견된 kinesin 유사 유전자의 homology 가 많은 motor domain을 이용하여 primer를 합성한 후 이를 이용하여 Schizosaccharomyces pombe 로부터 kinesin heavy chain 의 유전자를 클론하였다. 염기서열을 결정한 결과 2496 bp의 해독틀을 가지고 있으며 832개의 아미노산으로 이루어진 분자량이 96 kd의 kinesin heavy chain을 암호화하는 것이 밝혀졌다. 기존에 밝혀진 다른 organism 의 kinesin 과 서열을 비교한 결과 새로이 클론된 S.pombe 의 kinesin 은 C-terminal motor subfamily 에 속함이 밝혀졌다.

• 대한한의학회지
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• 제23권4호
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• pp.45-54
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• 2002

Objectives: In order to investigate the relationship of Radix Trichosanthis components and the melanin synthesis, the author has analyzed the cell viability and tyrosinase activity, melanin content and morphologic changes in n-hexane, EtOAc, n-BuOH, and H2O fraction. Methods: At first, in order to determine the concentration of the Radix Trichosanthis component, the author investigated the viability of B16 melanoma cell. To measure the effects of Trichosanthes kirilowii extracts (n-BuOH, n-Hexane, EtOAc, H2O fractions) on the viability of A549 cells, A549 cells were treated with various concentrations (from 0.5 to $25{\;}\mu\textrm{g}/ml$) of components of Trichosanthes kirilowii. After 24hrs, the cell viability was measured by MTT assay. The EtOAc components of Trichosanthes kirilowii decreased the viability of A549 cells in a dose-dependent manner. H2O and n-BuOH components had no cell toxicity till $25{\;}\mu\textrm{g}/ml$, the n-hexane component showed minor cell toxicity at $25{\;}\mu\textrm{g}/ml$ and the EtOAc component cell toxicity was revealed at $5{\;}\mu\textrm{g}/ml$ concentration. Results: 1. The results of tyrosinase activity and the Radix Trichosanthis component; n-hexane and EtOAc components controlled it effectively; the n-BuOH components were less effective. 2. The results of melanin content analysis showed that the n-hexane and EtOAc components effectively inhibited, the n-BuOH fraction inhibited less, and H2O component didn't inhibit the terminal melanin formation. 3. In the n-BuOH and H2O component there were no changes, but in the n-hexane component the melanin content was effectively inhibited. 4. In the EtOAc fraction, although the melanin content was inhibited, the cell count was evidently suppressed, Of all of the Radix Trichosanthis components, the n-Hexane and EtOAc fractions inhibited the melanin synthesis best, but owing to its toxicity, the EtOAc components inhibited the cell count. Conclusion: The above results demonstrated that Radix Trichosanthis n-hexane fraction efficiently inhibited the tyrosinase activity and melanin synthesis.