• Title/Summary/Keyword: phage

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Studies on the Genetic Recombination by Intraspecific Fusion of Lactobacillus casei Protoplast (Lactobacillus casei의 동종간 세포융합에 의한 유전자 재조합에 관한 연구)

  • Young Jin Baek;Hyeong Suk Bae;Young Kee Kim;Min Yoo;Hyun Uk Kim
    • Microbiology and Biotechnology Letters
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    • v.14 no.4
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    • pp.319-324
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    • 1986
  • After intraspecific fusion of Lactobacillus casei protoplasts, the recombinants have been studied for their lactose utilization, protease activity and phage resistance. L. casei C-M phenotypes constituted 46% of the fused cells when tested against phages, and L. casei 3-M phenotypes 42% of the fused cells, and 12% of the recombinants developed the resistance to both parent types of phages. The acid production and proteolytic activity of recombinants evidenced the similar trends. There was no difference in Hind III digests of plasmid DNA between parent cells and recombinants, but the reconlbinant cells were found to possess only one type of plasmid, either of 1. casei C-M or of L. casei 3-M.

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Circular Permutation of the DNA Genome of Temperate Bacteriophage $\PhiFC1$ from Enterococcus faecalis KBL 703

  • Kim, Young-Woo;Jang, Se-Hwan;Hong, Bum-Shik;Lim, Wang-Jin;Kim, Chan-Wha;Sung, Ha-Chin;Chang, Hyo-Ihl
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.457-463
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    • 1999
  • The physical map of bacteriophage $\PhiFC1$ DNA was constructed with the restriction endonucleases SalI, BamHI, EcoRI, XbaI, and AvaI. The 40.5-kb DNA restriction map is shown to be circularly permuted representing the headful packaging mechanism of the phage. The DNA restriction fragments containing the packaging initiation site(pac) was localized on the restriction map and the nucleotide sequences of the region were analyzed. Four open reading frames (ORFs), following one another with the same orientation, were found at the region. The 2nd ORF (ORF-ts) has significant amino acid sequence homologies to the previously known terminase small subunits of other bacteriophages. The putative terminase small subunit gene has a presumptive NTP-hydrolysis motif and a helix-turn-helix motif. The cleavage site for the first round of packaging was found to be located at the coding sequence of the putative terminase small subunit gene. The fourth ORF, even if partially sequenced, has a good amino acid sequence homology to the portal vertex proteins of other bacteriophages representing the evolutionarily conserved arrangements of genes near the pac site of this bacteriophage, $\PhiFC1$.

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Isolation and Characterization of Bacteriophages Against Pseudomonas syringae pv. actinidiae Causing Bacterial Canker Disease in Kiwifruit

  • Yu, Ji-Gang;Lim, Jeong-A;Song, Yu-Rim;Heu, Sunggi;Kim, Gyoung Hee;Koh, Young Jin;Oh, Chang-Sik
    • Journal of Microbiology and Biotechnology
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    • v.26 no.2
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    • pp.385-393
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    • 2016
  • Pseudomonas syringae pv. actinidiae causes bacterial canker disease in kiwifruit. Owing to the prohibition of agricultural antibiotic use in major kiwifruit-cultivating countries, alternative methods need to be developed to manage this disease. Bacteriophages are viruses that specifically infect target bacteria and have recently been reconsidered as potential biological control agents for bacterial pathogens owing to their specificity in terms of host range. In this study, we isolated bacteriophages against P. syringae pv. actinidiae from soils collected from kiwifruit orchards in Korea and selected seven bacteriophages for further characterization based on restriction enzyme digestion patterns of genomic DNA. Among the studied bacteriophages, two belong to the Myoviridae family and three belong to the Podoviridae family, based on morphology observed by transmission electron microscopy. The host range of the selected bacteriophages was confirmed using 18 strains of P. syringae pv. actinidiae, including the Psa2 and Psa3 groups, and some were also effective against other P. syringae pathovars. Lytic activity of the selected bacteriophages was sustained in vitro until 80 h, and their activity remained stable up to 50℃, at pH 11, and under UV-B light. These results indicate that the isolated bacteriophages are specific to P. syringae species and are resistant to various environmental factors, implying their potential use in control of bacterial canker disease in kiwifruits.

Widdrol Blocks 3T3-L1 Preadipocytes Growth and Differentiation Due to Inhibition of Mitotic Clonal Expansion

  • Yun, Hee-Jung;Kim, Jeong-Hwan;Jeong, Hyun-Young;Ji, Hyang-Hwa;Nam, Soo-Wan;Lee, Eun-Woo;Kim, Byung-Woo;Kwon, Hyun-Ju
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.806-813
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    • 2012
  • Adipocyte differentiation is strongly associated with obesity, which causes metabolic disorders. In this study, we investigated the inhibitory effects of widdrol on 3T3-L1 preadipocyte growth and differentiation. Widdrol decreased lipid droplet accumulation and down-regulated adipogenic transcription factors such as C/$EBP{\alpha}$, C/$EBP{\beta}$, and $PPAR{\gamma}$. Widdrol blocked preadipocyte proliferation and differentiation through the inhibition of mitotic clonal expansion, which was accompanied by the failure of degradation of p21, a cyclin-dependent kinase inhibitor. Cell-cycle analysis clearly indicated that widdrol actively induces cell-cycle arrest at the G1-S phage transition, causing cells to remain in the preadipocyte state. Moreover, widdrol increased p21 expression and inhibited Rb phosphorylation in preadipocyte incubated in a hormone medium. Therefore, these findings clearly suggest that widdrol blocks preadipocyte growth and differentiation through the inhibition of mitotic clonal expansion by p21-and Rb-dependent G1 arrest and can be developed as a potent anti-adipogenic agent for reducing obesity.

Rhizoma Scirpi induced Apoptosis in Human Cervical Carcinoma HeLa Cells (삼릉(三稜)이 자궁경부암세포(子宮頸部癌細胞)(HeLa cell)의 Apoptosis에 미치는 영향(影響))

  • Hong, Ki-Cheul;Kim, Joo-Yeon;Kong, Bok-Cheul;Choi, Chang-Min;Yoo, Sim-Keun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.18 no.4
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    • pp.10-23
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    • 2005
  • Purpose : This study is to examine the ability of Rhizoma Scirpi (RS) to induce HeLa cell viability. Methods : We culture HeLa cell which is human metrocarcinoma cell in D-MEM included 10% fetal bovine serum(Hyclone Laboratories) below $37^{\circ}C$, 5% CO2. Then we observed apoptosis of log phage cell which is changed cultivation liquid 24 Hours periodically. Results : 1. RS induces mitochondria membrane potential collapse. 2. P38 MAPK is involved in RS-induced death in HeLa cells. 3. P38 MAPK is involved in RS-induced apoptosis in HeLa cells. 4. P38 MAPK reguates RS-induced caspase-3, -8 and -9 activation in HeLa cells. 5. The inhibition of caspase regulates RS-induced cell death in HeLa cells. 6. RS induces mitochondria membrane potential collapse in HeLa cells. 7. P38 MPK is involved in the regulation of Bcl-2 and Bfu in HeLa cells.8. RS regulates the expression of Bcl-2 and Bax in HeLa cells. 9. SR induces p38 MAPK activation in HeLa cells. Conclusion : RS induces apoptosis in HeLa cells via p38 MAPK activation.

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Cloning and Sequencing Analysis of the Repressor Gene of Temperate Mycobacteriophage L1

  • Sau, Subrata;Chattoraj, Partho;Ganguly, Tridib;Lee, Chia Yen;Mandal, Nitai Chandra
    • BMB Reports
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    • v.37 no.2
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    • pp.254-259
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    • 2004
  • The wild-type and temperature-sensitive (ts) repressor genes were cloned from the temperate mycobacteriophage L1 and its mutant L1cIts391, respectively. A sequencing analysis revealed that the $131^{st}$ proline residue of the wild-type repressor was changed to leucine in the ts mutant repressor. The 100% identity that was discovered between the two DNA regions of phages L1 and L5, carrying the same sets of genes including their repressor genes, strengthened the speculation that L1 is a minor variant of phage L5 or vice versa. A comparative analysis of the repressor proteins of different mycobacteriophages suggests that the mycobacteriophage-specific repressor proteins constitute a new family of repressors, which were possibly evolved from a common ancestor. Alignment of the mycobacteriophage-specific repressor proteins showed at least 7 blocks (designated I-VII) that carried 3-8 identical amino acid residues. The amino acid residues of blocks V, VI, and some residues downstream to block VI are crucial for the function of the L1 (or L5) repressor. Blocks I and II possibly form the turn and helix 2 regions of the HTH motif of the repressor. Block IV in the L1 repressor is part of the most charged region encompassing amino acid residues 72-92, which flanks the putative N-terminal basic (residues 1-71) and C-terminal acidic (residues 93-183) domains of L1 repressor.

Putative Secondary Structure of Human Hepatitis B Viral X mRNA

  • Kim, Ha-Dong;Choi, Yoon-Chul;Lee, Bum-Yong;Junn, Eun-Sung;Ahn, Jeong-Keun;Kang, Chang-Won;Park, In-Won
    • BMB Reports
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    • v.28 no.6
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    • pp.509-514
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    • 1995
  • A putative secondary structure of the mRNA for the human hepatitis B virus (HBV) X gene is proposed based on not only chemical and enzymatic determination of its single- and double-stranded regions but also selection by the computer program MFOLD for energy minimum conformation under the constraints that the experimentally determined nucleotides were forced or prohibited to base pair. An RNA of 536 nucleotides including the 461-nucleotide HBV X mRNA sequence was synthesized in vitro by the phage T7 RNA polymerase transcription. The thermally renatured transcripts were subjected to chemical modifications with dimethylsulfate and kethoxal and enzymatic hydrolysis with single strand-specific RNase T1 and double strand-specific RNase V1, separately. The sites of modification and cleavage were detected by reverse transcriptase extension of 4 different primers. Many nucleotides could be assigned with high confidence, twenty in double-stranded and thirty-seven in Single-stranded regions. These nucleotides were forced and prohibited, respectively, to base pair in running the recursive RNA folding program MFOLD. The results suggest that 6 different regions (5 within X mRNA) of 14~23 nucleotides are Single-stranded. This putative structure provides a good working model and suggests potential target sites for antisense and ribozyme inhibitors and hybridization probes for the HBV X mRNA.

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A Design Study of Phase Changing Heat Exchanger for Environmental Control System (환경조절장치용 상변화열교환기의 개념설계연구)

  • Yoo, Young-June;Oh, Chang-Mook;Lee, Hyung-Joo;Min, Seong-Ki
    • Proceedings of the Korean Society of Propulsion Engineers Conference
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    • 2010.11a
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    • pp.628-635
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    • 2010
  • Properties of bleed air that is air source of ECS(Environmental Control System) can be rapidly changed with airplane engine operating conditions during flight. Therefore, ECS can be operated at a high performance or not during flight. So, high performance ACM has to be developed in order to flight safely. A adaptability of phase changing heat exchanger was esteemed at ACM type ECS in this study. As a result of this study, it is found that ECS outlet temperature can be controlled in a certain range with the phase changing phenomenon.

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Characterization of T7 RNA Polymerase Transcription Elongation Complex in Sequence-specific Transcription Termination (염기서열 특이적 전사종결부위에서 T7 RNA 중합효소 전사연장복합체 특성에 관한 연구)

  • Shin, Ji-Young;Lee, Sang-Soo
    • The Journal of Natural Sciences
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    • v.14 no.1
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    • pp.39-50
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    • 2004
  • T7 RNA polymerase is a single subunit RNA polymerase able to accomplish whole transcription process without auxiliary factors. In order to study transcription elongation mechanism of phage T7 RNA polymerse, stepwise walking of RNA polymerase was established by immobilizing biotinylated DNA template with streptavidin bead, series of active and stable elongation complexes were obtained, Transcripts were radio isotope labeled at the 16thm 17th and 18th nucleotide residues so stable elongation transcription complex of T7 RNA polymerase containing 22-40 nucleotide residues could be identified. We identified the positions of stablely formed transcription elongation complexes of termination site in intrinsic hairpin-independent PTH terminator sequence through the established stepwise walking of wild-type of mutant R173C T7 RNA polymerases. The results suggest that stable elongation transcription complexes were at the site of passing PTH terminator signal by mutant R173C RNA polymerase.

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An Immunological Approach to ABA Receptor and its Gene

  • Xie Zhou;Jin, Zhen-hua;Zheng, Zhi-fu;Kai Xia;Zhang, Neng-gang;Wan, Yin-sheng;Sang, Yong-ming;Chen, Kao-shan;Liu, Shi-ming
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.06a
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    • pp.68-78
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    • 1996
  • Two types of immunoloigcal probes, anti-ABBP Abs, have been developed. The purified ABBP from ABA-C1-BSA-sepharose 4B column was identified by PAGE and appeared in one band of about 56KD, as well as showed a specific binding ability and a high affinity for ABA (Kd2.0$\times$10-9 mol/L). Unexpectedly, the existence of rRNA with a length of around 300 nucleotides could be found, when the ABBP was digested with proteinase K and identified by eletrophorsis on an agarose gel (1%). As a result, about 120 cDNA clones coding maize 17s RNA and only one cDNA clone coding ABBP (24cDNA) were obtained from 200,000 seperated phage plaques by the anti-ABBP pAbs. 24cDNA had 1075bp and contained an open reading frame coding 254 amino acids. The anti-idiotypic Ab raised against an ABA MAb showed the ability of either mimicking ABA or competing with ABA. The localization of ABBPs in plant cell was investigated.

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