• Title/Summary/Keyword: pcbCD

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Cloning and Expression of pcbCD Genes in Escherichia coli from Pseudomonas sp. DJ-12 (Pseudomonas sp. DJ-12의 pcbCD 유전자의 클로닝과 Escherichia coli에서의 발현)

  • Kim, Chi-Kyung;Sung, Tae-Kyung;Nam, Jung-Hyun;Kim, Chang-Young;Lee, Jae-Koo
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.40-46
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    • 1994
  • The pcb genes of Pseudomonas sp. DJ-12 coded for the catabolism of polychlorinated biphenyl (PCBs) and biphenyl. The products of the pcbCD genes were 2,3-dihydroxy-4'-chlorobiphenyl dioxygenase and meta-cleavage product (MCP) hydrolase, which acted on degradation of 2,3-dihydroxy-4'-chlorobiphenyl to 4-chlorobenzoate. The pcbCD genes were cloned in E. coli XLl-Blue, and then the pcbD gene was further subcloned. As a metabolite transformed from 2,3-dihydroxybiphenyl by the cloned cell of E coli CU103, benzoate was detected by the resting cell assay. The enzyme activities of 2,3-dihydroxybiphenyl dioxygease and MCP hydrolase produced in the cloned cells E. coli CU103 and CU105 were about 17 and 3 times higher than those of Pseudomonas sp. DJ-12, respectively.

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Reidentification of Comamonas sp. Strain DJ-12 and Analysis of its pcbABC2D2 Genes Responsible for Degradation of 4-Chlorobiphenyl. (Comamonas sp. Strain DJ-12 의 재동정 및 4-Chlorobiphenyl 분해유전자 pcbABC2D2 의 분석)

  • 이준훈;박동우;강철희;채종찬;이동훈;김치경
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.121-126
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    • 2004
  • Comamonas sp. strain DJ-12 is a 4-chlobiphenyl(4CB)-degrading bacterium that was reidentified from Pseudomonas sp. DJ-12. The genomic DNA was isolated from the strain DJ-12 and amplified by PCR with primers for cloning pcbABCD genes responsible for degradation of 4CB. The amino acid sequences deduced from the nucleotide sequences of pcbA1, pcbA2, pcbA3, pcbA4, pcbB, pcbC2, and pcbD2 genes showed 91, 87, 99, 87, 97, 90 and 87% homologies with those of Pseudomonas sp. KKS102, respectively. The pcbC1D1 genes that are involved in the degradation of (4-chloro)1,2-dihydroxybiphenyl produced from 4CB by pcbAB gene products were previously reported in the recombinant plasmid pCU1 from Pseudomonas sp. DJ-12. However, the pcbC2D2 genes in the plasmid pCT4 and pCT5 cloned from Comamonas sp. DJ-12 in this study showed 51 and 62% homologies with those of pcbC1D1 in their nucleotide sequences. The pcbC1D1 and pcbC2D2 genes were found by Southern hybridization to be located at different loci on the chromosome of DJ-12 strain. These results indicate that Comamonas sp. strain DJ-12 has two different sets of pcbCD genes responsible for deg-radation of (4-chloro)1,2-dihydroxybiphenyl.

Cloning and Expression in E. coli of the HOPDA Hydrolase Gene from Pseudomonas sp. P20

  • Lim, Jong Chul;Chae, Jong Chan;Kim Youngsoo;Kim, Hyong Bai;Kim Chi Kyung
    • Journal of Microbiology
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    • v.34 no.4
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    • pp.349-354
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    • 1996
  • Pseudomonas sp. P20 is a natural isolate which is capable of degrading biphenyl and 4-chlorobiphenyl. From a clone of pCK1022 harboring pcbCD genes of Pseudomonas sp P20, a pcbD gene encoding 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoic acid (HOPDA) hydrolase was subcloned in Escherichia coli XL-1-Blue by using pBluescript SK(+) vector. The 2.8-kb HindII fragment harboring the pcbD gene cloned in pCK 1024 had a single site for each of XhoI, SalI, BstXI, and XbaI restriction enzymes. Escherichia coli CK1024 had a single site for each of XhoI, SalI, BstXI, and XbaI restriction enzymes. Escherichia coli CK1024 carrying pCK0124 degraded HOPDA to benzoate and 2-hydroxypenta-2, 4-dienoate by HOPDA hydrolase encoded by pcbD gene as effectively as E coli CK 1022 HARBORING pcbCD genes.

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Effect of Porcine Cancellous Bones on Regeneration in Rats with Calvarial Defect (랫드의 두개골 결손부에서 돼지 해면질골이 골재생에 미치는 영향)

  • Yoo, Kyeong-Hoon;Kim, Se-Eun;Shim, Kyung-Mi;Park, Hyun-Jeong;Choi, Seok-Hwa;Kang, Seong-Soo
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1207-1213
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    • 2010
  • The purpose of this study was to evaluate the effect of porcine cancellous bone as a scaffold in a rat calvarial defect model. Critical-sized defects were created in 30 male Sprague-Dawley rats. The animals were divided into critical defect (CD, n=10), $\beta$-tricalcium phosphate (TCP) graft (BT, n=10) and porcine cancellous bone graft (PCB, n=10) groups. Each defect was filled with $\beta$-TCP mixed with fibrin glue or porcine cancellous bone powder mixed with fibrin glue. In the CD group, the defect was left empty. All rats were sacrificed at 8 weeks after bone graft surgery, and bone formation was evaluated by gross observation, plain radiography, micro-computed tomography scanning and histological evaluation. Repair of bone defect was the least in the CD group, and significant new bone formation was observed in the PCB group. Grafting of porcine cancellous bone was more efficient for regenerating new bone than grafting $\beta$-TCP.

Cloning of pcb Genes in Pseudomonas sp.P20 Specifying Degradation of 4-Clorobiphenyl (4-Chlorobiphenyl을 분해하는 Pseudomonas sp. P20의 pcb 유전자군의 클로닝)

  • 남정현;김치경
    • Microbiology and Biotechnology Letters
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    • v.22 no.4
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    • pp.353-359
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    • 1994
  • Pseudomonas sp. P20 was a bacterial isolate which has the ability to degrade 4-chlorobi- phenyl(4CB) to 4-chlorobenzoic acid via the process of meta-cleavage. The recombinant plasmid pCK1 was constructed by insetting the 14-kb EcoRI fragment of the chromosomal DNA containing the 4CB-degrading genes into the vector pBluescript SK(+). Subsequently, E. coli XL1-Blue was transformed with the hybrid plasmid producing the recombinant E. coli CK1. The recombinant cells degraded 4CB and 2,3-dihydroxybiphenyl(2,3-DHBP) by the pcbAB and pcbCD gene products, respectively. The pcbC gene was expressed most abundantly at the late exponential phase in E. coli CK1 as well as in Pseudomonas sp. P20, and the level of the pcbC gene product, 2,3-DHBP dioxygenase, expressed in E. coli CK1 was about two-times higher than in Pseudomonas sp. P20. The activities of 2,3-DHBP dioxygenase on catechol and 3-methylcatechol were about 26 to 31% of its activity on 2,3-DHBP, but the enzyme did not reveal any activities on 4-methylcatechol and 4-chlorocatechol.

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pKT230 벡터를 이용한 Pseudomonas sp. P20의 2,3-Dihydroxybiphenyl Dioxygenase 유전자의 클로닝

  • Kim, Ji-Young;Kim, Chi-Kyung;Ka, Jong-Ok;Min, Kyung-Hee;Park, Yong-Keun
    • Microbiology and Biotechnology Letters
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    • v.24 no.6
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    • pp.657-663
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    • 1996
  • Pseudomonas sp. P20 isolated from the polluted environment is capable of degrading biphenyl and 4-chlorobiphenyl. The pcbABCD genes responsible for degradation of biphenyl and 4-chlorobiphenyl were cloned using pBluescript SK(+) from the chromosomal DNA of Pseudomonas sp. P20 to construct pCK1 and pCK102, harbouring pcbABCD and pcbCD, respectively. The 2, 3-DHBP dioxygenase gene, pcbC, was cloned again from pCK102 by using pKT230 which is known as a shuttle vector and pKK1 hybrid plasmid was constructed. The E. coli KK1 transformant obtained by transforming the pKK1 into E. coli XL1-Blue showed 2, 3-DHBP dioxygenase activity. The specific 2, 3-DHBP dioxygenase activity of E. coli KK1 was similar to that of the E. coli CK102, but much higher than those of the natural isolates, Pseudomonas sp. DJ-12 and Pseudomonas sp. P20.

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The Survey about Toxic Material Contamination at the Streamwater around the Sudokwon Kandfill Area (수도권 매립지 주변 하천의 독성물질 오염실태 조사)

  • Kang, Chang-Min;Lee, In-Hyun;Jang, Won
    • Journal of Environmental Science International
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    • v.7 no.4
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    • pp.565-572
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    • 1998
  • This study estimate the degree of contamination in the streamwater around the Sudokwon landfill site. It was sampled at 10 sites in Jan., Apr., Jul. and Oct., 1996. There were analyzed five kinds of toxic chemical material-CN, AES, PCB, As, Org-p, and four kinds of heavy metal-Pb, Hg, Cd, $C^{+6}$ The result are 1)The COD was generally increased to compare before landfall, 2) The Org-P and PCB were not detected at all points, 3) The concentrations of Pb, $C^{+6}$ and As were lower than the environmental criteria values, 4) The CN, Hg and Cd were over envirommental criteria values. and so emergent regulation is needed, 5) The effects of the streamwater contamination were not only the leachate of the landfall, but also the small factories and agricultural land around the landfill.

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A design of HomePNA2.0 PHY. (10Mbps급 HomePNA2.0 PHY. 회로 설계)

  • 박성희;구기종;김종원
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.27 no.12C
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    • pp.1282-1287
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    • 2002
  • In this Paper, we present the design of 10Mbps HomePNA(Home Phoneline Networking Alliance) PHY which is Home Network Technology using phone-line. It is connected with external interface through MII(Media Independent Interface) and AFE(Analog Front End) Interface. 10Mbps HomePNA PHY is composed with Management Block IEEE 802.3 CSMA/CD MAC(Media Access Control) Block Modulator block and Demodulator block. For their verification, we designed a prototype FPGA PCB board using XPC860T made in Motorola. We verified HomePNA frame data transmission using a driver program based Linux kernel. we verified rate negotiation by HomepNA 2.0 Link Layer Protocol.

Assessment of Environmental Pollution for Streams of Andong City in Gyeongbuk Province Using Invertebrate Biomarker and Chemical Residual Analysis (무척추동물 생체지표와 화학잔류량 분석을 통한 경북 안동지역내 하천들의 환경오염 평가)

  • Ryoo Keon-Sang;Choi Jong-Ha;Kim Young-Gyun;Cho Sung-Hwan;Lee Hwa-Sung
    • Journal of Environmental Science International
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    • v.14 no.6
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    • pp.583-596
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    • 2005
  • Samples of water, soil, and sediment were taken from 10 streams of Andong city in Gyeongbuk province in October 2004. To assess the degree of environmental pollution for each stream site, the chemical analyses of pollutants such as T-N, T-P, COD, heavy metal, organophosphorous and organochlorine pesticides, and dioxin-like PCB congeners were implemented using the standard process tests or the U. S. EPA methods. In addition, biological assessment using insect immune biomarkers was conducted on the same environmental samples to complement the chemical assessment. Except Waya stream (T-N; 2.91 mg/L, T-P; 0.16 mg/L, COD; 14.0 mg/L) with above the environmental quality standards, the T-P and COD concentrations of 9 sites are relatively low. The contents of Pb and Cd in samples taken from each stream were much lower than environmental quality standards. However, in comparison with soil samples of other streams, several times higher concentrations of Pb and Cd were found in locations at Mi, Gilan, Yeonha, and Waya stream sites. Dementon-S-methyl, diazinon, parathion, and phenthoate compounds among organophosphorous pesticides were detected as concentrations of ppb levels, respectively, from soil samples collected in the vicinity of Gilan, Mi, Norim, and Waya stream. On the other hand, 16 organochlorine pesticides and 12 dioxin-like PCB congeners selected in this study were not found in all samples. In particular, considering significant disrupting effects of Waya stream's samples on insect immune capacity, this stream seems to be contaminated with investigated and/or univestigated pollutants in this study.