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Molecular Cloning and Expression of $\beta$-Xylosidase Gene from Thermophilic Alkalophilic Bacillus sp. K-17 into Escheyichia cozi and Bacillus subtilis (고온, 호알칼리성 Bacillus속 K-17 균주의 $\beta$-Xylosidase유전자의 Escherichia coli 및 Bacillus subtilis의 클로닝 및 발현)

  • Sung, Nack-Kie;Chun, Hyo-Kon;Chung, Duck-Hwa;Shim, Ki-Hwan;Kang, In-Soo
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.436-439
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    • 1989
  • The chromosomal DNA fragments of thermophilic alkalophilic Bacillus sp, K-17, a potent xylanhydrolyzing bacterium, were ligated to a vector plasmid pBR322 and transformed into Escherichia coli HB101. The plasmid pAX278, isolated from a transformant forming yellow color on the LB agar plate containing 1 mM p-nitrophenyl- $\beta$-xylopyranoside, was found to enable the transformants to produce p-xylosidase. The 5.0 kilobase insert of pAX278 had single sites for EcoRI, PstI, XbaI, and PvuII, and 2 sites for BglII. Biotinylated pAX218 was hybridized to 0.9 kb as well as 5.0 kb fragment from Bacillus sp. K-17 DNA on nitrocellulose filter. pGX718 was constructed by inserting the 5.0 kb HindIII fragment of pGX278 at the HindIII site of pGR71, E. coli and B. subtilis shuttle vector. The enzymatic properties of $\beta$-xylosidase from E. coli HB101 carrying recombinant plasmid were the same those of $\beta$-xylosidase from Bacillus sp. K-17.

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Molecular Cloning of Human Genomic DNA for Epinephrine Synthesizing Enzyme, Phenylethanolamine N-Methyltransferase (Epinephrine 합성효소인 phenylethanolamine N-methyltransferase의 인간 genomic DNA의 유전자 크로닝)

  • Suh, Yoo-Hun;Huh, Sung-Oh;Chun, Yang-Sook;Kim, Hun-Sik;Lim, Jung-Kyoo;Park, Chan-Woong
    • The Korean Journal of Pharmacology
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    • v.24 no.1
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    • pp.1-10
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    • 1988
  • To obtain information about the structure of the human phenylethanolamin N-methyltransferase (PNMT) and to further define the extent of the evolutionary relationships among PNMT molecules of several spesies, a full length cDNA clone for bovine adrenal PNMT was used to screen a charon 4A genomic library. One phage was isolated and identified, which included the entire PNMT gene. The length of inserted genomic DNA was 13.1-Kilobase (Kb) containing two internal EcoRI sites. Construction of a restriction map and subsequent Southern and dot blot analysis with 5'-and3'-specific cDNA probes allowed the identification of exon-containing fragments. This is the first report of the cloning of gene for human epinephrine synthesizing enzyme.

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Preparation and Quality Characteristics of Namul-kim bugak Using Aster yomena (쑥부쟁이를 활용한 나물김부각 제조 및 품질특성)

  • Lee, Gyeong-Eun;Park, Su-Jin;Jung, Yeon-Kwon;Go, Geun-Bae;Son, Byeong-Gil;Jeong, Ji-Suk
    • Korean journal of food and cookery science
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    • v.32 no.1
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    • pp.50-57
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    • 2016
  • In this study, kim (dried laver) bugak ("KB"), Aster yomena bugak ("SB"), and namul (vegetables)-kim bugak (combination of Aster yomena and kim) ("NB") were prepared and their quality characteristics were investigated. NB was made by slicing Aster yomena, seasoning it with perilla seeds, attaching it to kim and applying glutinous rice paste, followed by freeze-drying. SB had a higher oil absorption rate than KB, reflecting the high oil absorption rate of namul (here, Aster yomena). DPPH radical scavenging activity was highest for SB; whereas, NB had higher DPPH radical scavenging activity than KB, indicative of the higher antioxidant capacity of namul, as compared to kim. With longer storage period, moisture content tended to be increased, while maximum load, yield stress and hardness tended to be decreased. Overall preference was in the order of NB>KB>SB, without statistical significance. Acid value and peroxide value of NB, stored at $25^{\circ}C$ for 40 days, tended to be increased gradually but did not meet the criteria for rancidity. Therefore, NB could be stored at $<25^{\circ}C$ for > 40 days. Furthermore, it would be possible to make functional bugak using a variety of namul, as well as Aster yomena.

An Effective Multimedia Data Transmission in Ad-Hoc Networks Based on Bluetooth (블루투스를 이용한 애드혹 네트워크에서의 효율적인 멀티미디어 데이터 전송)

  • Kim, Byoung-Kug;Hong, Sung-Hwa;Hur, Kyeong;Eom, Doo-Seop
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.33 no.3B
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    • pp.112-122
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    • 2008
  • Basing on Piconet, The Bluetooth System forms network and transmits data. There is one Master and maximum 7 Slave bluetooth devices in one piconet. A job scheduler performed by Master bluetooth device, gives the chance of data transmission to Slave bluetooth devices, which connected to Master, using polling method in piconet. The maximum data rate is 723.2 kb/s when it uses ACL link with DH5 packet type in a piconet which is constructed by two bluetooth devices. However, if there are one master and two slave devices in a piconet, then the maximum data rate is reduced to a half(361.6kb/s), because a master device has to support same data rate for all connected devices. And, there is the defect in scatternet when data transmission rate becomes low(Maximum rate: 302.2kb/s). This paper proposals the new ad-hoc network topology called "DoublePico"for overcome the low data transmission in scatternet which is constructed by piconets. The method of doublepico that represented in this paper makes high data transfer rate(Maximum rate: 457.57kb/s) in bluetooth ad-hoc networks.

Effect of Estrogen on the Gestational Profiles in Gene Expression of Placental Lactogen I, II and Pit-i in the Rat Placenta (흰쥐 태반에서 Placental Lactogen I과 II 그리고 Pit-1의 유전자 발현에 미치는 에스트로겐의 영향)

  • 정진권;강성구;강해묵;이병주
    • The Korean Journal of Zoology
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    • v.39 no.1
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    • pp.115-121
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    • 1996
  • To investigate gestational profiles in gene expression of placental lactogen I fpL4), PL-lI and Pit-1, RNA samples were extracted from the placentas of pregnant day 12 to 20 at 2 day intervals. Northern blots showed changes in gene expression of PL4, - 11 and Pit-i. Sizes of PL-l and -II mRNA were changed and amounts of PL-I, -H and Pit-1 mRNA increased during progress of gestation. To examine the effect of estrogen on the gene expression of PL-I, -Il and Pit-1, pregnant female rats were ovariectomized (OVX) and daily injected with estradiol (OVX + E). OVX markedly lowered the amount of PL4 and 41 mRNA, and shifted niRNA size from 1 kb to i 3 kb in PL-l mRNA and 0.6 kb to i kb in PL-ll mRNA, respectively. OVX had no effect on the mRNA size of Pit-1, but markedly attenuated Pit-1 mRNA level. Estrogen injection reversed the effect of OVX on the size-shift but not on the amount of PL4 and -Il mRNA. Replacement of E partially recovered OVX-induced inhibition of Pit-i mRNA level. Present results suggest that estrogen may play a pivotal role on the gene expression of PL-l and -Il such as alternative RNA splicing and/or polyadenylation, and Pit-1 may be involved in the gene expression of PL-l and 41 by estrogen.

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Molecular cloning of casein gane which is expressed in mammary glands (유선조직에서 특이적으로 발현되는 카제인 유전자의 클로닝(I))

  • Choe, In-Ho;Bae, Bong-Jin;Lee, Chang-Su
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.21 no.1
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    • pp.53-66
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    • 1995
  • The gene for ${\gamma}$-casein, a milk protein, is a member of a family of casein gene which is expressed in mammary glands of the animal during the late gestation and lactation periods binder the influence of various hormones. In order to elucidate tile mechanisms b)'which hormones regulate the coordinate induction of milk protein genes, the mouse ${\gamma}$-casein gene was isolated and characterized. The ${\gamma}$-casein gene was screened from a mouse genomic library constructed in bacteriophage EMBL3 with the ${\gamma}$-casein CDNA used as probe and one clone was obtained. The ${\gamma}$-casein CDNA as probe was partially sequenced and contained ATG start codon and 5'-noncoding region. The cloned genomic DNA was digested with Sal I restriction enzyme, by which the insert DNA can be isolated from EMBL3 vector. Three DNA bands were observed and the size of DNAs was approximately 28kb, 14kb and 9Kb, respectively Accordingly the size of the insert DNA was calculated with approximately 23Kb. The result of Southern blot analysis, however, showed that the cloned genomic DNA was not hybridized with the synthetic oligonucleotides (40 mer) of cDNA 5'-end region, but it was hybridized with the y -casein CDNA. This means that tile cloned y -casein gene may not contain its promoter region. The ${\gamma}$ -casein genomic DNA containing the promoter region has been screening from mouse genomic library with oligonucleotides of CDNA 5'-end region as probe, and twenty-nine clones was obtained.

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Molecular Cloning and Expression of a Cellulolytic Xylanase Gene from Bacillus circulans in Escherichia coli (Bacillus circulans 기원의 Cellulolytic Xylanase 유전자의 대장균에서의 클로닝 및 발현)

  • 이동석;김지연;김한복
    • Korean Journal of Microbiology
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    • v.36 no.3
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    • pp.196-202
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    • 2000
  • A gene for cellulolytic xylanase of Bacillus circulnns ATCC21365 was cloned on pUC 19 in Eschwichia coli. The recombinant plasniid pXLI80 contained an 1.8 id, inselt composed of0.5 kb and 1.3 kb PslI fragments derived from B, circulans. The 0.5 kh fragment in the upstream region of 1.3 kb one was confirmed lo be indispensable for not only expression but also hyperexpression of the cloned gene. The transformant overproduced the xylanase 135 times greater than that produced by the orlginal B circulnns. The optimum pH and temperature of the cloned enzyme we]-e pH 5.2 and $60^{\circ}C$, respectively. Heal pretl-eatment at TEX>$55^{\circ}C$C for 1 Indid not cause inhibition of the activity of this enzyme. The elm.ynie could hydl-olyre CMC and lichenan as well as xylan to produce xylose(or GI), xylohiose(or G2) and xylolnose(or G3) as inah products. Hence We defined the cloned enzyme as a cellulolytic xylanase. The SDS-PAG electrophoretic mobility and zyiiogram of this enzyme derived from whole cell extracts or c~~lture supematants or E. coli(pXL180) indicated a molecular weight of 45,000 and nonprocessing of the enzyme in the peilplasln of E. coli.

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High Prevalence and Genotypic Characterization of Metallo-β-Lactamase (MBL)-Producing Acinetobacter spp. Isolates Disseminated in a Korean Hospital (국내 대학병원에서 분리된 Metallo-β-Lactamase (MBL) 생성 Acinetobacter spp. 분리주의 높은 출현율과 유전형 특징)

  • Yum, Jong Hwa
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.4
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    • pp.444-452
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    • 2019
  • Carbapenem resistance, mediated by the major acquired metallo-β-lactamase (MBL) genes, has been increasingly reported, particularly for clinical isolates of Acinetobacter spp. Of the 191 nonduplicate clinical isolates of the carbapenem-nonsusceptible Acinetobacter spp. evaluated, 125 isolates (65.4%) were positive for the modified imipenem or meropenem-Hodge test, and 49 isolates (25.7%) were positive for the imipenem-EDTA+SMA double disk synergy test (DDS). PCR and sequencing of the blaVIM-2-allele and blaIMP-1-allele showed that 29 A. baumannii isolates and 1 A. calcoaceticus isolate had blaVIM-2, whereas 16 A. baumannii isolates and 2 A. calcoaceticus isolates had blaIMP-6; 1 isolate of the A. genomospecies 3 had blaVIM-2 and blaAIM-1. All the above MBL genes belong to class 1 integron. The size of class 1 integron encompassing blaVIM-2 or blaIMP-6 ranges from 2.8 kb to 3.2 kb in clinical isolates of A. baumannii, and 3.2 kb to 3.5 kb in clinical isolates of A. genomospecies 3. blaVIM-2 was most often located first or second in the class 1 integron, and these integrons often included aacA4. Due to dispersion of the MBL-producing Acinetobacter spp. as well as integron, which may encompass various resistance genes, there is an expectation for the increase of multidrug resistant Gram-negative bacteria, including resistance of carbapenems such as imipenem or meropenem. Hence, the development of new antimicrobial agents for treating severe Acinetobacter spp. infections is needed.

Zircon Morphology and Petrochemistry of Mesozoic Plutonic rocks in Seonsan Area, Korea (선산 지역 중생대 심성암류의 저어콘 헝태 및 암석화학)

  • 이윤종;박순자;장용성;정원우;김중욱;황상구;윤성효
    • The Journal of the Petrological Society of Korea
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    • v.13 no.2
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    • pp.81-102
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    • 2004
  • The plutonic rocks in Seonsan area are divided into dioritic-syenitic rock, gneissose granite, biotite granite and fine grained biotite granite. These rocks intruded into the Pre-cambrian metamorphic complex and are all covered by the Cretaceous Nakdong formation. According to modal minerals, dioritic-syenitic rock corresponds to quartz monzonite, granodiorite, tonalite fields, whereas all the other plutonic rocks fall in granite field. Petrochemically the dioritic-syenitic rock is lower in SiO$_2$ content, differentiation index and Larsen index than all the other plutonic rocks. About the zircon morphology, dioritic-syenitic rock shows (100) dominant type but other granitic rocks exhibit mixed types between (100) and (110) type. The dioritic-syenitic rock could be crystallized in higher temperature than the other plutonic rocks. The plutonic rocks correspond to calc-alkaline rock series, and belong to I-type granite and mostly magnetite-series in magmatic origin. In plutonic processes, the dioritic-syenitic rock with 5kb vapor pressure could intrude into the metamorphic batement at 17km deep below the surface. Later the gneissose granite with lower 3kb vapor pressure could intrude at 10km deep. Sequentially the biotite granite with 0.7kb could intrude at 2km deep. Finally the fine grained biotite granite with 3kb vapor pressure could intrude at 10km deep.

BCR (Brown Color Repressor) gene isolation related to mycelial browning of Lentinus edodes (표고균사 갈변과 관련된 BCR (Brown Color Repressor) 유전자 분리)

  • Kim, Young-Ho;Park, Soo-Cheol;Jhune, Chang-Sung;You, Chang-Hyun;Sung, Jae-Mo;Kong, Won-Sik
    • Journal of Mushroom
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    • v.10 no.3
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    • pp.120-128
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    • 2012
  • Recently sawdust cultivation of Shiitake mushroom (Lentinula edodes) is increasing. It is important to make mycelia to be brown on the substrate surface. This browned surface in sawdust cultivation plays an important role like as artificial bark of the oak log, which protects the other pests and suppresses water evaporation in the substrate. In order to isolate genes which related to brown color formation, differential display method was used. Two cDNA fragments obtained by DD-PCR were 1.2 and 1.6kb and these were expressed in white colored mycelia from L. edodes, but not brown colored mycelia. Partial sequencing of these cDNA fragments showed that the 1.6kb cDNA had 100% identity with the microsatellites gene from Dugenia polichroa. However, the other 1.2kb cDNA fragment had poly T tail on 3' region of partial open reading frame on 5' region. The new primer designed based on the sequence of 1.2kb cDNA was constructed. RT-PCR analysis using the newly designed 0.12kb cDNA specific primer showed that the gene was only expressed in white color mycelia, but not in brown color mycelia. Sequence analysis of 5' region of this 1.2kb cDNA revealed that this gene contained partial open reading frame consisted of 110 amino acid. Homology search using DNASIS database showed that this gene had high sequence homology of 66.7% in DNA level and 69.2 % in amino acid level with dTDP-glucose 4,6-dehydratases gene from Arabidopsis thaliata. The dTDP-glucose 4,6-dehydratases gene was known to be function to have tolerance with oxidation stress. These results strongly suggest that this gene isolated from white mycelia of L. edodes might have a function of repressor against mycelia browning. Therefore I designated this gene as BCR (Brown Color Repressor) gene.