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Cloning of a Paenibacillus sp. Endo-${\circ}$-1,4-Glucanase Gene and Its Coexpression with the Endomyces fibuliger ${\circ}$-Glucosidase Gene in Saccharomyces cerevisiae

  • KIM, HYUNJIN;JI-YOUNG YANG;HYEON-GYU LEE;JAEHO, CHA
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.685-692
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    • 2001
  • A gene, Egl, from Paenibacillus sp. KCTC 8848P encoding endo-${\circ}$-1,4-glucanase was cloned and expressed in Escherichia coli. It consisted of an open reading frame of 1,191 bp for a protein that consisted of 397 amino acids with a molecular weight of 44,539 Da. The deduced amino acid sequence of the endo-${\circ}$-1,4-glucanase gene had a 94% similarity to the endo-$\beta$-1,4-glucanase of Bacillus polymyxa. The Egl gene was also expressed in Saccharomyces cerevisiae secreting Endomyces fibuliger $\beta$-glucosidase (BGL1) under the control of the alcohol dehydrogenase (ADC1) gene promoter, S. cerevisiae transformant producing both endo-${\circ}$-1,4-glucanase and ${\circ}$-glucosidase grew on carboxymethyl cellulose as the sole carbon source.

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Nucleotide Sequence Analysis of an Endo-Xylanase Gene (xynA) from Bacillus stearothermophilus

  • Cho, Ssang-Goo;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.5 no.3
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    • pp.117-124
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    • 1995
  • A gene (xynA) encoding the endo-xylanase (E.C.3.2.1.8) from Bacillus stearothermophilus was cloned in E. coli, and its complete nucleotide sequence was determined. The xynA gene consists of a 636 base pairs open reading frame coding for a protein of 212 amino acids with a deduced molecular weight of 23, 283 Da. A putative signal sequence of 27 amino acid residues shows the features comparable with the Bacillus signal sequences; namely, the signal contains a positively charged region close to the N-terminus followed by a long hydrophobic string. The coding sequence is preceded by a possible ribosome binding site with a free energy value of -16.6 kcal/mol and the transcription initiation signals are located further upstream. The translation termination codon (TAA) at the 3 end of the coding sequence is followed by two palindrome sequences, one of which is thought to act as a terminator. The xynA gene has a high GC content, especially in the wobble position of codons (64%). Comparison of the primary protein sequence with those of other xylanases shows a high homology to the xylanases belonging to family G.

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Identification and Expression Analyses of Equine Endogenous Retroviruses in Horses

  • Gim, Jeong-An;Kim, Heui-Soo
    • Molecules and Cells
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    • v.40 no.10
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    • pp.796-804
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    • 2017
  • Endogenous retroviruses (ERVs) have been integrated into vertebrate genomes and have momentously affected host organisms. Horses (Equus caballus) have been domesticated and selected for elite racing ability over centuries. ERVs played an important role in the evolutionary diversification of the horse genome. In the present study, we identified six equine ERV families (EqERVs-E1, I1, M2, P1, S1, and Y4), their full-length viral open reading frames (ORFs), and elucidated their phylogenetic relationships. The divergence time of EqERV families assuming an evolutionary rate of 0.2%/Myr indicated that EqERV-S3 (75.4 million years ago; mya) on chromosome 10 is an old EqERV family and EqERV-P5 (1.2 Mya) on chromosome 12 is a young member. During the evolutionary diversification of horses, the EqERV-I family diverged 1.7 Mya to 38.7 Mya. Reverse transcription quantitative real-time PCR (RT-qPCR) amplification of EqERV pol genes showed greater expression in the cerebellum of the Jeju horse than the Thoroughbred horse. These results could contribute further dynamic studies for horse genome in relation to EqERV gene function.

Molecular identification and expression analysis of a natural killer enhancing factor-A from black rockfish Sebastes schlegelii

  • Lee, Jeong-Ho;Kim, Joo-Won;Park, Chan-Il
    • Journal of fish pathology
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    • v.22 no.3
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    • pp.343-352
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    • 2009
  • Natural-killer-cell-enhancing factor (NKEF) belongs to the newly defined peroxiredoxin (Prx) family. It was originally isolated from human erythroid cells. The black rockfish NKEF cDNA was identified through the expressed sequence tag (EST) analysis of PBLs libraries. The full-length NKEF cDNA was 1433 bp long and contained an open reading frame (ORF) of 594 bp that encoded 198 amino-acid residues. The 5' UTR had a length of 39 bp, and the 3’UTR 800 bp. The deduced amino-acid sequence of the black rockfish had a density 93.4, 92.9, 87.8, 85.8, 84.8, 83.8, 80.3, 79.7, 77.2, and 75.2% that of the pufferfish, olive flounder, channel catfish, zebrafish, chicken, common carp, Myotis lucifugus, cattle, human PrxI, rat PrxI, human NKEF-A, and Xenopus tropicalis, respectively. The NKEF gene was expressed in all the tissues of the black rockfish. The RT-PCR indicated that the NKEF transcripts were predominantly in the spleen and gill, less dominantly in the PBLs, head kidney, trunk kidney, and liver, and least in the intestine and muscles. This is the first report on the existence of the NKEF-A gene in black rockfish.

Function of mORF1 Protein as a Terminal Recognition Factor for the Linear Mitochondrial Plasmid pMLP1 from Pleurotus ostreatus

  • Kim, Eun-Kyoung;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.37 no.4
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    • pp.229-233
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    • 1999
  • The mitochondrial plasmid pMLP1 from a white-rot fungus, Pleurotus ostreatus, is a double-stranded DNA containing 381 bp terminal inverted repeat (TIR) whose 5'-ends are covalently bound by terminal proteins. The plasmid contains two major open reading frames (ORFs), encoding putative DNA and RNA polymerases, and a minor ORF encoding a small, highly basic protein. To identify the DNA binding activity that recognizes the TIR region of pMLP1, gel retardation assays were performed with mitochondrial extracts. A specific protein binding to a region between 123 and 248 nt within TIR was observed. We examined whether the gene product of mORF1 bindes to this region specifically. E. coli cell extract which contains an overproduced mORF1 protein formed a complex specific to the region between 123 and 248 nt. Inclusion of mORF1 protein in the specific complex formed between P. ostreatus mitochondrial extract and TIR was confirmed by a supershift assay using polyclonal antibodies against the mORF1 protein. Our result suggest that the product of mORF1 may function as a terminal region recognition factor (TRF), recognizing an internal region in TIR.

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Complete genome sequence analysis Hosta virus X and comparison to other potexviruses

  • Park, M.H.;K.H. Ryu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.113.1-113
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    • 2003
  • A potexvirus, Hosta virus X (HVX-Kr), causing mosaic and mottle symptoms was isolated from hosta plants (Hosta spp.), and its entire genome RNA sequence was determined. in Korea using cDNA library and RACE methods. The genome of HVX encodes five open reading frames coding for viral replicase, triple gene block (TGB), and viral coat protein (CP) from the 5'to 3' ends, which is a typical genome structure of potexviruses. The 3-terminal region of the virus includes the TGBI (26 kDa), TGB2 (13 kDa), TGB3 (8 kDa), and 23 kDa coat protein (CP) and the 3-nontranslated region (NTR). The CP gene of the type isolate of HVX (HVX-U) was amplified by RT-PCR and its nucleotide sequence was determined. The CPs of HVX-Kr and HVX-U had 100% and 98.9% identical amino acids and nucleotides, respectively. Most of the regions of the genome HVX had over 50% nucleotide identical to other sequenced potexviruses. This is the first report of complete genome sequence information of HVX and molecular evidence supporting the virus as a distinct species of the genus Potexvirus.

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Complete nucleotide sequence of genome RNA of Daphe virus S and its relationship n the genus Carlavirus (oral)

  • Lee, B.Y.;K.H. Ryu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.115.2-116
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    • 2003
  • Complete genomic nucleotide sequence of Daphe virus S (DVS), a member of the genus Carlavirus, causing leaf distortion and chlorotic spot disease symptoms in daphne plants, has been determined in this study. The genome of DVS contained six open reading fames coding for long viral replicase, triple gene block, 36 kDa viral coat protein (CP) and 12 kDa from the 5' to 3' ends, which is a typical genome structure of carlaviruses. Two Korean isolates of DVS isolates were 98.1% and 93.6% amino acid identical in the CP and 12kDa, respectively. The CP gene of DVS shares 25.2-55.2% and 42.9-56.1% similarities with that of 19 other carlaviruses at the amino acid and nucleotide levels, respectively. The 3'-proximal 12 kDa gene of DVS shares 20.2-57.8% amino acid identities with that of 18 other members of the genus. The 3' noncoding region of DVS consists of 73 nucleotides with long excluding poly A tract, and shares 69.1-77.1% identities to the known carlaviruses. In the phylogenetic analyses of the two proteins, DVS was closely related to Helenium virus S and Chrysanthemum virus B. This is the first complete sequence information for the DVS, and further confirms the classification of DVS as a distinct species of the genus Carlavirus.

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Interdisciplinary and philosophy of Diderot Considered through De l'interprétation de la nature ('융·복합'과 디드로의 철학 - 『자연해석론』을 중심으로 -)

  • PAEK, Chan-Wook
    • Cross-Cultural Studies
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    • v.33
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    • pp.143-169
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    • 2013
  • Recently, because of the danger and damages that following the steady development of scientific technologies, natural science is faced with many humane and ethical problems. So it is asserted the interdisciplinary with social and human science. However, it comes commonly that the ethical issues such as world view and one's view of life caused the development of sociology, especially development of the market economy. But in many cases, the interdisciplinary can be useful for widen the view of scholars. The interdisciplinary is actually connected with the problems of philosophy, and located in that domain. In this case, this is noticed as a model to the philosophers in 18th century, especially Denis Diderot. Diderot published De $l^{\prime}interpr{\acute{e}}tation$ de la nature during editing the Encyclopedia, from there he picked out the contents from piles of documents of Encyclopedia. Even though the contents or opinions of De $l^{\prime}interpr{\acute{e}}tation$ de la nature are inaccuracy or erroneous, it shows that how human-social science and natural science encountered. Diderot studied mathematics and then Diderot accepted to the natural science proposal, he approaches philosophy with translate English books to the French. Next he understood natural science by reading Buffon and Maupertuis, and during working for Encyclopedia, he possessed his knowledge that he can claim his opinion to other scholars. However in this De $l^{\prime}interpr{\acute{e}}tation$ de la nature, Diderot who sometimes rebutted other scholars' theory and demonstration, tried to build a philosophy on metaphysics in order to it was important for himself that he imposed the methods of science and importance of experience. Anyhow, this De $l^{\prime}interpr{\acute{e}}tation$ de la nature cause consider the recognition of Diderot in the field of natural science, and is suggested as a model about his Nature. This mean that it is an expression of his philosophy, and the content is found from natural philosophy and empirical philosophy. Like giving these attache the importance of method study for science and technique, these are targeted the promotion of popularization of natural-science and scientific-technology. Also it advocates fulfilling from reasonable philosophy to empirical philosophy. Therefore, the philosophy which was speculative and abstracted became his philosophy which was writing the meaning, as waiting the discovery of science. And at that time, the humanities made interdisciplinary with natural science.

cDNA Cloning and Stage-Dependant Expression of Arylphorin Gene from Chinese Oak Silkworm, Antheraea pernyi (작잠(Antheraea pernyi) 아릴포린(Arylphorin) 유전자의 cDNA 클로닝 및 아릴포린 유전자의 발육시기 의존성 발현양상)

  • Lee, Sang-Mong;Hwang, Jae-Sam;Park, Nam-Sook;Kim, Yong-Gyun;Kim, Keun-Ki;Son, Hong-Joo;Park, Hyun-Chul;Jin, Byung-Rae
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1193-1200
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    • 2010
  • The cDNA cloning and developmental profiles of the mRNA for A. pernyi arylphorin was determined. The complete A. pernyi arylphorin cDNA sequence comprised 2,234 bp (without the poly $A^+$ tail), including an open reading frame of 2,112 bp beginning with a methionine ATG at bp34. The A. pernyi arylphorin contained 704 amino acids which are highly enriched in aromatic amino acids, phenylalanine and tyrosine. The calculated molecular mass of the A. pernyi arylphorin from the ORF was 83,439 Da. The deduced amino acid sequence of A. pernyi arylphorin showed 78, 71, 62 and 64% identity with those of H. cecropia, M. sexta $\alpha$ subunit, M. sexta $\beta$ subunit and B. mori storage protein. In Northern blot analysis, the A. pernyi arylphorin mRNA only in the fat body of the 5th instar larvae was responsible for gene expression of the protein, and the synthetic activity of the mRNA was detected strongly in the early larvae, but not in the middle or late-stage larvae. In addition, a very weak signal in mRNA activity was detected in pupal stages, but this was considered to be inactive mRNA after reviewing the results of the labeling experiment of this protein.

The Evaluation of Thermal Aging for Insulation Paper with Mineral and Vegetable Oil (광유 및 식물성 절연유에서 절연지의 열적 열화 특성 평가)

  • Lee, Byung-Sung;Song, Il-Keun;Kim, Dong-Myung;Han, Sang-Ok
    • Proceedings of the KIEE Conference
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    • 2004.07c
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    • pp.1558-1560
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    • 2004
  • Liquid-immersed pole transformers have utilized an insulation system consisting of natural cellulose-based materials and mineral oil for many years. If the transformers are installed with the abruptly changing load over limitation, its insulation materials are apt to unexpected aging with high temperature. Therefore we have been conscious of developing a transformer that is more thermally stable then conventional types. To measure temperature distribution in the winding of conventional and newly designed transformers, temperature sensors were installed fordirect reading of oil and conductor temperatures at transformers. And we conducted accelerating aging test for these transformers. The measuring data had made use of the design to hybrid insulation system. Also we could find the possibility of this insulation system to apply in pole mounted transformers.

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