• Title/Summary/Keyword: KC101

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Modigication of host cells and Expression of Recombinant E. coli trp plasmids for the increased Production of Tryptophan in Klebsiella pneumoniae (Klebsiella pneumoniae에서 트립토판 생산증대를 위한 숙주개발 및 재조합 trp plasmid의 발현)

  • 지연태;홍광원;박장현;이세영
    • Korean Journal of Microbiology
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    • v.25 no.1
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    • pp.46-51
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    • 1987
  • In order to increase the production of tryptophan by maximizing expression of recombinant trp plasmid, Klebsiella pneumoniae KC 105(pheA tyrA trpE trpR tyrR) was genetically modified. KC 107, inosine monophospate(IMP) auxotroph from KC 105 and KC 108, histidine(His) auxotroph from KC 107 were also derived respectively to increase phosphoribosylpyrophosphate(PRPP) production which is required for tryptophan biosynthesis. From KC 107 phosphoribosylpyrophosphate consumption which is required for tryptophan biosynthesis. From KC 107 and KC 108, KC 109 and KC 110, both arginine auxotrophs were derived respectively. To investigate the expression of recombinant trp plasmid in the selected K. pneumoniae mutants, the auxotrophic mutants were transformed with recombinant trp plasmids pSC 101-$trpE^{FBR}$, pSC 101-trpL(.DELTA.att) $trpE^{FBR}$ (pSC 101-trp-AF). Amount of tryptophan produced and activities of tryptophan synthase of $trpR^{-}$ mutant (KC 100) and $tyrR^{-}$ mutnat(KC 105) containing recombinant plasmid pSC 101-trp operon were increased by 30-40% as compared with KC 99(pheA tyrA trpE) containing recombinant plasmid pSC 101-trp operon. Activities of tryptophan synthase and production of tryptophan of KC 108 ($His^{-}$) and KC 109($Arg^{-}$) containing recombinant plasmid pSC 101-trp operon were increase by two-fold as compared with KC 107 containing pSC 101-trp operon.

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Effect of Hwalhyulsungjang-san and KC101 Composed of Oriental Medicinal Stuffs on Physical Development in Growing Rats (활혈성장산과 KC101이 흰쥐의 성장발육에 미치는 영향)

  • 박승만;한찬규
    • The Journal of Korean Medicine
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    • v.24 no.1
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    • pp.1-8
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    • 2003
  • Purpose : Nowadays, there are many studies of growth retardation treatment by oriental medical therapy. This study was performed to evaluate the effect of Hwalhyulsungjang-san (Huoxuechengzhang-san) and KC101 on physical development in growth, Methods : 60 young (3 weeks old) male rats, whose mean weight was $46.8{\pm}0,7g$, were divided into 5 groups : groups A, B, C, and D were fed an experimental diet containing respectively Hwalhyulsungjang-san 2.5%, Hwalhyulsungjang-san 7.5%, KC101 5.0%, and KC101 10.0%, and the other group were fed an AIN-diet containing neither Hwalhyulsungjang-san nor KC101 as a control. Study of each group used 2 rats and was repeated in 6 times. Body weight, daily weight gain, dietary intake, body length, femur length, backbone length and serum concentration of IGF-I and hemoglobin were measured. Result : 1. Body weight and daily weight gain of group D (KC101 10%) were highest; dietary intake of group D was also increased by 13% over the control group. 2. Average body length of group D was increased to 1.69cm, which was 3.2% over the control group. Backbone length of group D was also increased 4% compared to control group. 3. Serum concentration of IGF-I of group D increased 17% compared to control group. 4. Serum concentration of hemoglobin of group B (Hwalhyulsungjang-san 7.5%) increased 8% compared to control group. Conclusion : This study showed that Hwalhyulsungjang-san and KC101 composed of oriental medicinal stuffs have effect on physical development in growing rats. There were not any statistical meaning compared to control group, but this study suggests that intake of these compounds from a young age may lead to increase physical development in growth.

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Cloning and Expression of the Bacillus thruingiensis var. kurstaki HD-1 Crystal Protein gene in Eschelichia coli

  • Sang Hyn Kim;You
    • Journal of Sericultural and Entomological Science
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    • v.35 no.2
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    • pp.129-133
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    • 1993
  • The 44Md plasmid of Bacillus thruingiensis var. kurstaki HD-1(B. t k HD-1) was partially digested with Sau3AI and the fragments were cloned into E. coli HB101 on vector pBR322. Of 2, 950 clones with a recombinant pBR322, only one clone KC1 was determined to have the gene for crystal toxic proteins from the 44Md plasmid of B. t k HD-1 at the BamHI site of pBR322. The recombinant pBR322 was named pKC1 and its molecular size was 12kb. The KC1 produced a protein which was toxic to the silkworm and antigenically similar to the crystal toxic protein of B. t k HD-1. Also, electrophoretic mobility of the KC1 protein was apparently the same as that of the crystal toxic protein of B. t k HD-1.

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Breeding of Korean Ginseng (Panax ginseng C. A. Meyer)

  • Kwang-Tae Choi;Woo
    • Proceedings of the Ginseng society Conference
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    • 1998.06a
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    • pp.96-102
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    • 1998
  • A lot of individual ginseng plants were selected in the farmer's fields to develop new ginseng varieties with desirable traits. Among them, a promising line has been developed through comparative cultivation of several lines selected with pure line separation of local raced. It was then designated as "KC (denotes Korean Ginseng)" and tested in the regional yield and adaptation trials for 10 years. KG lines grew vigorously after 4 years of age. Especially, KG 102 line arson운 them showed traits of multiple and short stems. As for the root characters, the length of taproot of KG 101 line was longer than that of local race, Jakyung-jong, or other lines ailed the root weight of KG 102 line waIn 15 % higher than that of local race. In general, KG 101, KG 103, KG 104, and Hwangsauk-jong had good root shape. Total amount of ginsenosides of ginseng taproot was the highest in KG 103 line than in local race and other lines. In these studies, we, elected three superior lines, KG 101, KG 102, and KG 103, having characteristic of good root shape, high yield, and large amount of ginsenoside, respectively.

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Effect of Polygoni Multiflori Radix Water Extract on the Proinflammatory Mediators in RAW 264.7 Cells Induced by LPS (하수오(何首烏) 물추출물이 LPS로 유발된 RAW 264.7 Cells의 염증인자에 미치는 영향)

  • Jeong, Seong-Yong;Han, Hyo-Sang;Lee, Young-Jong
    • The Korea Journal of Herbology
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    • v.28 no.6
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    • pp.101-109
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    • 2013
  • Objectives : The purpose of this study was to investigate the effects of Polygoni Multiflori Radix Water Extract (PM) on the production of inflammatory mediators in RAW 264.7 mouse macrophages induced by lipopolysaccharide (LPS). Method : We examined effect of PM Extract on the cell viability of RAW 264.7 cells. Futhermore, we investigated anti-inflammatory effect of PM Extract by the production of proinflammatory cytokines such as NO, intracellular calcium, interleukin(IL)-$1{\alpha}$, IL-3, IL-$1{\beta}$, IL-6, interferon inducible protein-10(IP-10), keratinocyte-derived chemokine(KC) and vascular endothelial growth factor(VEGF). Result : No significant changes have been found in the mouse macrophge cell viability by the PM Extract at the concentration of 25, 50, 100 and $200{\mu}g/mL$. The water extract of PM significantly inhibited the production of NO and intracellular calcium in the LPS-induced macrophages at the concentration of 25, 50, 100 and $200{\mu}g/mL$. The water extract of PM significantly inhibited the production of IL-$1{\alpha}$, IL-${\beta}$, IL-3, IP-10, KC, VEGF in the LPS-induced macrophages at the concentration of 50, 100, and $200{\mu}g/mL$; IL-6 at the concentration of 100 and $200{\mu}g/mL$ ; and IL-17 at $200{\mu}g/mL$. Conclusion : The water extract of PM significantly inhibited the production of NO, intracellular calcium, IL-$1{\alpha}$, IL-3, IL-${\beta}$, IP-10, KC, VEGF at the concentration of 50 ㎍/mL or higher in the LPS-induced macrophages with no changes in the cell viability of them. These results suggest that water extract of Polygoni Multiflori Radix has anti-inflammatory effect regulating the production of proinflammatory cytokines in the LPS-induced macrophages.

Phylogenetic Analysis of Korean Black Cattle Based on the Mitochondrial Cytochrome b Gene (mtDNA cytochrome b에 기초한 한국흑우의 계통유전학적 분석)

  • Kim, Jae-Hwan;Byun, Mi Jung;Kim, Myung-Jick;Suh, Sang Won;Kim, Young-Sin;Ko, Yeoung-Gyu;Kim, Sung Woo;Jung, Kyoung-Sub;Kim, Dong-Hun;Choi, Seong-Bok
    • Journal of Life Science
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    • v.23 no.1
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    • pp.24-30
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    • 2013
  • The purpose of this study was to identify genetic polymorphisms of the mitochondrial cytochrome b (mtDNA cyt b) gene in Korean black (KB) cattle breed and to analyze the genetic relationship between the KB and other breeds. We determined the complete sequence of the mtDNA cyt b gene in 38 KB cattle. We also analyzed their genetic diversity, and phylogenetic analysis was performed by comparison with Korean cattle (KC, called Hanwoo) and breeds from China and Japan. A nucleotide substitution was detected in the KB cattle, and two haplotypes were defined. In the neighbor-joining (NJ) tree, the haplotypes of KB were located in Bos taurus lineage with those of KC, Japanese black (JB), Yanbian and Zaosheng breeds. However, the haplotypes of Chinese breeds, excluding Yanbian and Zaosheng, were separated into B. taurus and B. indicus lineages. In the NJ tree of breeds based on Dxy genetic distances, Chinese breeds mixed with B. taurus and B. indicus lineages were located between B. indicus and B. taurus lineages. KB was contained within B. taurus lineage and was determined to be genetically more closely related to two Chinese (Yanbian and Zaosheng) breeds than to KC and JB. The haplotype distribution and the results of the phylogenetic analysis suggest that KB and KC have genetic differences in their mtDNA cyt b gene sequences.

Identification and Analysis of Putative Polyhydroxyalkanoate Synthase (PhaC) in Pseudomonas fluorescens

  • Lim, Ju Hyoung;Rhie, Ho-Gun;Kim, Jeong Nam
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1133-1140
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    • 2018
  • Pseudomonas fluorescens KLR101 was found to be capable of producing polyhydroxyalkanoate (PHA) using various sugars and fatty acids with carbon numbers ranging from 2 to 6. The PHA granules consisted mainly of a poly(3-hydroxybutyrate) homopolymer and/or poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer. Genomic DNA of P. fluorescens was fractionated and cloned into a lambda library, in which a 5.8-kb fragment that hybridized to a heterologous phaC probe from Ralstonia eutropha was identified. In vivo expression in Klebsiella aerogenes KC2671 (pUMS), restriction mapping, Southern hybridization experiments, and sequencing data revealed that PHA biosynthesis by P. fluorescens relied upon a polypeptide encoded by a 1,683-bp non-operonal ORF, which was preceded by a possible -24/-12 promoter and highly similar to DNA sequences of a gene encoding PHA synthase in the genus Pseudomonas. In vivo expression of the putative PHA synthase gene ($phaC_{Pf}$) in a recombinant Escherichia coli strain was investigated by using glucose and decanoate as substrates. E. coli (${phaC_{Pf}}^+$, pUMS) grown in medium containing glucose accumulated PHA granules consisting mainly of 3-hydroxybutyrate, whereas only a trace amount of 3-hydroxydecanoate was detected from an E. coli fadR mutant (${phaC_{Pf}}^+$) grown in medium containing decanoate. In vitro enzymatic assessment experiments showed that 3-hydroxybutyryl-CoA was efficiently used as a substrate of purified $PhaC_{Pf}$, suggesting that the putative PHA synthase of P. fluorescens utilizes mainly short-chain-length PHA precursors as a substrate.

유전공학적으로 변형시킨 R-plasmid 들의 전이에 미치는 균주와 pH 의 영향

  • 김희태;이성기;김치경
    • Korean Journal of Microbiology
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    • v.30 no.2
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    • pp.88-95
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    • 1992
  • The genetically engineered microorganisms (GEMS) could be released accidentally or ii)rexperimental purposes, as the genetic engineering, technique ha:. become very popular inany laboratories of biological sciences. But there have been littlt: informations on transkrbehavior of the genetically ~nodified genes in the natural en\ironmentx. In this stutly.antibiotic resistant bacteria were isolated from nat.ural waters. and then GEM strains wereconstructed i'rom the natural isolate (NI) by ~noclification oi' the Km' plasmitl. Thetransferability of the plasmids in the GEM and NI strains were examinetl by con-jugationin Luria-Bertani broth :it 30$^{\circ}$C. Also the cff'ccts 01' mating strain and pH on their transferfrequency and rearrangement of the plasmids in tl-~ec o~ijugantsM ere comp:irati\ely stuclictl.I'hc transkr frequency of Km' plasmid in donor of GEM and N1 strains wah similar a.;about 10 ' when co~ljugation was conducted wit11 M'I'I strain is recipient at pH 7. butthat of 1)KCOOI was lowered to 1.2X 10 '. And when the lab. stlain was uhccl as recipient.the transfer tendency of the plasmid was about same in both (;EM and NI strains usedas donor. All thc tionor 5trains. except for I)KC601. showecl the Ilighcs~ frequency of about10 ' at pH 7 and the frequcncics were lowered at both pH 5 and 9. Hut the mocliliedKm' plasmid in the cloned strain of DKC601 was transferred hy very low frequency of10 "at pH 5 ant1 7 comparing to other GEM strains. especiall! any co~~.jugantws ere notobtained at pH 4 and 9 even after conjugation for 6 hours. Rearrangement of the plasmidstranskrred into the lab. strain was not found in the conjugants. I\ulcornerut a lot of rearrangclncntwas ohservecl nlhen they were transferred into the NI strain. Such a rearrangement wasmore severe when donor was GEM strain rather then NI strain Hut such ;r phenomenonwas less affected by p!-l values.r phenomenon was less affected by p!-l values.

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Study on Relaxing Effect of Oxybutynin on the Contractile Response of Arterial Smooth Muscle (동맥근 수축에 대학 Oxybutynin의 이완효과에 관한 연구)

  • Ko, Jae-Ki;Baik, Yung-Hong
    • The Korean Journal of Pharmacology
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    • v.24 no.1
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    • pp.93-101
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    • 1988
  • Pharmacological actions of an antispasmodic agent, oxybutynin were investigated in the isolated procine coronary arteries. The coronary rings were contracted by acetylcholine (ACh) and KCl in a dose-dependent fashion. The ACh-induced contractions were signifcantly potentiated by removal of endothelium and $EC_{50}=0.52\;{\mu}M$ of intact endothelial rings was about 2 times greater than $EC_{50}=0.28\;{\mu}M$ of rings without the endothelium. These results suggest that the endothelium plays an inhibitory role in ACh-induced contraction. Oxybutynin and atropine inhibited dose-dependently $1.0\;{\mu}M$ ACh-induced contraction and atropine inhibited dose-dependently $1.0\;{\mu}M$ ACh-induced contraction and the $IC_{50s}$ were 11.0 nM and 0.47 nM, respectively. Atropine did not affect 35 mM KCl-induced contraction but oxybutynin inhibited the contraction to the basal tension in a dose-dependent manner. The $IC_{50}$ of oxybutynin on the KCl-induced contraction was $49.7\;{\mu}M$. The dose-response curve to ACh was parallelly shifted to the right by pretreating coronary rings with $IC_{50}$ of atropine (0.47 nM) or oxybutynin (11.0 nM) but the curve to KC1 was rightward shifted in a noncompetitive manner under pretreatment with $IC_{50}$ of oxybutynin $(49.7\;{\mu}M$). Oxybutynin inhibited $0.1\;{\mu}M$ Bay K 8644-induced contraction to the basal tension in a dose dependent manner, but $35\;{\mu}M$ histamine-induced contraction was inhibited to only 50e/e of the original level even in maximal concentration $(5{\times}10^{-4}M)$ of oxybutynin. These results suggest that oxybutynin causes antispasmodic action through sensitive blocking action on muscarinic receptors and inhibitory action on calcium influx in the procine coronary artery.

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