• Title/Summary/Keyword: EcoR I

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Molecular Cloning of $\beta$-Galactosidase Gene from Neisseria lactamica 2118 into Escherichia coli MC 1061 (Neisseria lactamica 2118의 $\beta$-galactosidase 유전자의 대장균으로의 클로닝)

  • Lee, Jong-Su
    • The Journal of Natural Sciences
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    • v.5 no.1
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    • pp.37-45
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    • 1992
  • The gene coding for $\beta$-galactosidase of Neisseria lactamica 2118 was cloned into Escherichia coli MC 1061. The isolated 6.5 Kb EcoR I fragement and 7.2 Kb BamH I fragment of chromosomal DNA in Southern hybridization were ligated to a vector plasmid pBR322 and then transformed into Escherichia coli MC 1061 cells. Finally, I obtained three clones as $\beta$-galactosidase positive clone by colony hybridization and Southern hybridization($\beta$-galactosidase probe: lac Z gene of pMC1871). Three recombinant plasmids(pNL.13. 17 and 24) were found to contain the 7.2Kb BamH I fragment originated from Neisseria lactamica 2118 chromosomal DNA by Southern hybridization and pNL 24 was showed high homology to probe especially and also its physical map was constructed.

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Development of Controlled Gas Nitriding Furnace : Controlled Gas Nitriding Technology and Present Situation in Korea (질화포텐셜 제어 가스질화로 개발(I) : 제어질화 및 국내 기술 현황)

  • Won-Beom Lee;Sukwon Son
    • Journal of the Korean Society for Heat Treatment
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    • v.36 no.1
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    • pp.40-46
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    • 2023
  • Controlled nitriding is a technology that controls the nitriding potential based on the gas partial pressure received through an IOT-based sensor. Controlled nitriding is characterized by easy control of the phase of the nitride compound and excellent reproducibility of quality. In particular, it is possible to form a compound layer of excellent quality with fewer pores on the surface. However, despite these advantages, the application of controlled nitriding still needs to be improved in Korea. This paper explains the characteristics of controlled nitriding and describes the future direction and the problems of controlled nitriding in Korea.

Restriction endonuclease maps of three plasmids from bacillus thuringiensis serovar israelensis 4Q1 (Bacillus thuringiensis serovar israelensis 4Q1로부터 분리된 plasmid 제한효소지도 작성)

  • Faust, R.M.;;;C.L.Meyers-Dowling;P.E.McCawley
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.122-128
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    • 1985
  • Bacillus thuringiensis serovar israelensis 4Q1 contains 8 different covalently closed-circular (CCC) plasmids of molecular weight 204, 267, 109, 103, 16, 7.6, 6.4, and 5.0kb. The three smallest plasmids, designated pBti6, and pBti8 may prove to be useful as cloning vectors because of thier size and ease of isolation. The three plasmids were incubated separately with 9 different restriction enzymes and 7 of the enzymes tested cleaved one or more of the plasmids. Plasmid pBti6 has a single site for Bg1 II, Pst I and Pvu II, two sites for Bc1 I and Eco RI, and five sites for Hind III. Plasmid pBti7 has a single site for Bam HI and Pst I, two sites for Hind III, and three sites for PvuII. Plasmic pBti8 has a single site for Bam HI, BelI and Hind III, two sites for Eco RI, and three sites for Bgl II and Pvu II. Composite restriction enzyme maps for pBti6, pBti7 and pBti8 were constructed. The sites of restriction enzyme cleavage were determined by single, double and partial digests of the plasmid DNA. All the restriction sites were aligned relative to the single Bgl II(pBti6), Pst I(pBti7) or Hind III(pBti8) site, respectively.

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Cloning and Characterization of Autonomously Replicating Sequence(ARS) from Kluyveromyces fragilis

  • HONG, SOON-DUCK;JONG-GUK KIM;TAKUYA NAGAMATSU;JOO-HYUN NAM;DONG-SUN LEE;SANG-YONG LEE;SUN-HWA HA
    • Journal of Microbiology and Biotechnology
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    • v.3 no.1
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    • pp.6-11
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    • 1993
  • An autonomously replicating sequence (Kf-ARS1) of Kluyveromyces fragilis was cloned from the genomic library which was constructed using pHN134 as a cloning vector to make a new host-vector system for the production of heterologous protein from K. fragilis as a host. The cloning vector pHN134 was composed of $Km^r, Ap^r$ and multiple cloning site in LacZ . A clone carrying Kf-ARS1 was isolated and the recombinant plasmid was designated as pIKD102. The cloned fragment was 2.3 kb (EcoRI/EcoRI) in length. Subcloning experiment showed that the region for ARS activity was 1.5 kb (SalI/EcoRI) fragment. It was shown that the Kf-ARS1 was active in Saccharomyces cerevisiae and Kluyveromyces fragilis.

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Marine Algal Flora and Ecological Role of Eisenia bicyclis in Dokdo, East Sea, Korea (한국 동해안 독도의 해조상 및 대황(Eisenia bicyclis) 부착기의 생물상)

  • Park, Seo Kyoung;Lee, Jung Rok;Heo, Jin Suk;An, Dae Sung;Lee, Haeng Pil;Choi, Han Gil
    • Korean Journal of Environment and Ecology
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    • v.28 no.6
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    • pp.613-626
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    • 2014
  • Seaweeds were identified after qualitative sampling at 14 stations of Dokdo coasts from May to July 2013 and seaweeds and animals inhabiting 23 Eisenia bicyclis plants were examined to evaluate its ecological role. Biomass was calculated by using regression between stipe length and weight of E. bicyclis. A total of 128 species were identified, including 18 green, 35 brown, and 75 red algae. Coarsely branched form was dominant functional group occupying 47.66% and 91 species (71.09%) were in ESG I group, growing in stable environmental coast. Also, R/P, C/P, and (R+C)/P were 2.14, 0.51, and 2.66, respectively showing temperate and mixed flora. Biomass and density of E.bicyclis were $23.74kg\;m^{-2}$ and 64 fronds $m^{-2}$. Twelve seaweeds and 83 animal species (15 Annelida, 25 Mollusca, 34 Arthropoda, 3 Echinodermata, and 6 others) were observed from 23 holdfasts and Ericthonius pugnax was dominant taxon having 538 (43.11%) of 1,248 animal individuals. These results indicate that E.bicyclis is a keystone species showing very important ecological role. In conclusion, the number of seaweeds increased because of intensive research and dominance of coarsely branched form and ESG I group seaweeds, representing that environmental condition of Dokdo is still intact.

Effects of Mitochondrial DNA Polymorphism on Growth Traits of Hanwoo (mt DNA 다형이 한우 성장에 미치는 영향)

  • Jeon, G.J.;Chung, H.Y.;Choi, J.G.;Lee, M.S.;Chung, Y.H.;Lee, C.W.;Park, J.J.;Ha, J.M.;Lee, H.K.;Na, K.J.
    • Journal of Embryo Transfer
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    • v.18 no.3
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    • pp.227-235
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    • 2003
  • 한우의 mt DNA cytochrome oxidase subunit I, II, 및 III complex지역의 유전적 다형현상을 제한효소를 이용하여 검출하였다. PCR primer 6종에 대하여 20가지 제한효소를 처리하였으며, Pst I, Pvu II, Rsa I, Eco RI, Bgl II, and Msp I 제한효소를 사용하여 유전적 변이를 검출하였다. 검출된 변이체와 한우의 성장과의 관련성을 조사한 결과 cytochrome oxidase subunit III complex 지역의 유전염기서열을 근거로 제작한 primer Mt9 좌위에서 제한효소 PvuII를 이용한 절단형과 체중형질 인 WT15(P<0.05) 및 WT18(P<0.01)에서 고도의 유의성이 관찰되었다. 아울러 , Mt9-Pvu II(P=0.07), Mt6-Bgl II(P=0.05), and Mt8-Rsa I(P=0.05) 좌위 또한 WT9, WTl5, and WT15에서 각각 통계적 유의성이 관찰되었다. 따라서 본 결과는 cytochrome oxidase subunit III complex segments가 candidate gene으로서 기초적 유전정 보 제공은 물론 유전적 개량을 위해 사용될 수 있을 것으로 사료된다.

Recycling of the Waste Rock and Tailings from Yangyang Iron Mine (양양철광산 선광 부산물의 순환자원화)

  • Jung, Moon Young;An, Yong Hyeon;Kim, Young Hun
    • Resources Recycling
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    • v.25 no.4
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    • pp.23-31
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    • 2016
  • It was found that there was no problem in recycling by-products (waste rock and tailings) from Yangyang iron mine themselves through matter conversion because they are not hazardous according to results of KSLT method. In case of using tailings as sub-materials of cement, it recommended the use of less than 3% tailings dosage not to exceed 0.6% of total alkali ($R_2O$) content based on standard quality of portland cement (KS L 5201). Non sintered eco-brick corresponding to class 1 quality of recycled clay brick (KS I 3013) can replace 15% of cement with tailings and 100% of general fine aggregate with waste rock from iron mine. As mentioned above, recycling the by-products (waste rock and tailings) as sub-materials of cement and non sintered eco-brick could gain both environmental and economic benefits, that is, reduction of scale and maintenance cost of tailing ponds, decrease of energy use and $CO_2$ emission.

Comparative Genetic Characterization of Plasmids of Agrobacterium Species Isolated in Korea (한국산 Agrobacterium plasmid의 유전학적 성상에 관한 연구)

  • Kim, Jung-Hye;Koo, Yong-Bum;Lee, Ki-Yung;Chung, Jae-Kyu
    • Journal of Yeungnam Medical Science
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    • v.1 no.1
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    • pp.41-48
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    • 1984
  • The soil bacterium Agrobacterium tumefaciens is a plant pathogen that cause3 crown gall tumors by infecting the wounded dicotyledonous plants and subsequent integration of bacterial DNA into plant nuclear DNA. Virulent A. tumefaciens strains harbor a large Ti (tumor-inducing) plasmid that carries genes essential for tumorigenesis. In the present study, 13 strains (Malus pumila Mill; $A_{1-3}$, Populus monilifera; $W_{1-6}$, Populus tomentiglandlosa; $P_{1-3}$ and Rosa species; $R_1$) of Agrobacterium isolated in korean crown gall tumors and plasmids were observed in 6 strains ($W_2$, $W_3$, $W_6$, $P_1$, $P_3$ and $A_2$). The test for crown gall tumor formation was resulted only in ATCC15955 and $KW_2$ strains inoculated into the stem of sun flower and the development was observed for 4 and 6 weeks after inoculation. Above two Ti plasmids (pTi) were purified by cesium chloride-ethidium bromide density gradient centrifugation and digested with restriction enzyme and fragments of pTiATCC 15955 and $pTiKW_2$ observed by EcoR I ; 25&27, Hind III; 23&21, BamH I ; each 20 and Hpa I ; 12&27, and sizes of pTiATCC15955 and and $pTiKW_2$ calculated as 200 and 87 kbases. Octopine was isolated from tumor tissue ($W_{1-6}$ and $P_{1-3}$) and these strains confirmed as octopine type.

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Cloning and Organization of the Ribosomal RNA Genes of the Mushroom Trichloma matsutake

  • Hwang, Seon-Kap;Kim, Jong-Guk
    • Journal of Microbiology and Biotechnology
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    • v.5 no.4
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    • pp.194-199
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    • 1995
  • A portion (7.4 kb) of ribosomal DNA tandem repeat unit from a genome of the mushroom T. matsutake has been cloned. A 1.75 kb EcoRI fragment was cloned first using S. cerevisiae 255 rRNA gene as a probe, and this was then used for further cloning. A chromosomal walking experiment was carried out and the upstream region of the 1.75 kb fragment was cloned using SmaI/BamHI enzyme, the size was estimated to be 5.2 kb in length. Part of the downstream region of the 1.75 kb fragment was also cloned using XbaI/BamHI enzymes. Restriction enzyme maps of three cloned DNA fragments were constructed. Northern hybridization, using total RNA of T. matsutake, and the restriction fragments of three cloned DNAs as probes, revealed that all four ribosomal RNA genes (large subunit[LSU], small subunit [SSU], 5.85 and 5S rRNA genes) are present in the cloned region. The gene organization of the rDNA are regarded as an intergenic spacer [IGS]2 (partial) - SSU rRNA - internal transcribed spacer [ITS]1 - 5.8S rRNA - ITS2 - LSU rRNA - IGS1 -5S rRNA - IG52 (partial).

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Phage Particle Proteins and Genomic Characterization of the Lactobacillus plantarum Bacteriophage SC 921. (Lactobacillus plantarum Bacteriophage SC 921의 phage particle protein 및 genome의 특성)

  • 김재원;신영재;심영섭;유승구;윤성식
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.117-121
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    • 1998
  • Bacteriophage SC 921 of Lactobacillus plantarum, isolated from kimchi, showed high lytic effects at 0.2 M.O.I. level. The phage particle contained 4 major proteins (48, 34, 32, 29 kDa). Intact DNA of phage SC 921 is a double stranded linear molecule, and the genomic size is approximately 66.5 kilobase pairs (kbp). Restriction analysis of the genome showed that Sma I gave single site cut and Xba I gave 2 site cuts, while Cla I, Kpn I, and EcoR I formed 4, 5, and 6 cuts, respectively. Hind III digested phage DNA to many fragments. A restriction map of genomic DNA was constructed using the restriction endonuclease Kpn I, Sma I, and Xba I. Bacteriophage SC 921 was compared with B2 phage which had been reported to infect Lactobacillus plantarum ATCC 8014(KCCM l1322). Bacteriophage SC 921 differs from B2 phage at least in thr size of its genome and phage particle proteins.

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