• 제목/요약/키워드: stable cells

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Construction of Conjugative Gene Transfer System Between E. coli and Moderately Thermophilic, Extremely Acidophilic Acidithiobacillus caldus MTH-04

  • Liu, Xianggmei;Lin, Jianqun;Zhang, Zheng;Bian, Jiang;Zhao, Qing;Liu, Ying;Lin, Jianqiang;Yan, Wangming
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.162-167
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    • 2007
  • A genetic transfer system for introducing foreign genes to biomining microorganisms is urgently needed. Thus, a conjugative gene transfer system was investigated for a moderately thermophilic, extremely acidophilic biomining bacterium, Acidithiobacillus caldus MTH-04. The broad-hostrange IncP plasmids RP4 and R68.45 were transferred directly into A. caldus MTH-04 from Escherichia coli by conjugation at relatively high frequencies. Additionally the broad-hostrange IncQ plasmids pJRD215, pVLT33, and pVLT35 were also transferred into A. caldus MTH-04 with the help of plasmid RP4 or strains with plasmid RP4 integrated into their chromosome, such as E. coli SM10. The $Km^r\;and\;Sm^r$ selectable markers from these plasmids were successfully expressed in A. caldus MTH-04. Futhermore, the IncP and IncQ plasmids were transferred back into E. coli cells from A. caldus MTH-04, thereby confirming the initial transfer of these plasmids from E. coli to A. caldus MTH-04. All the IncP and IncQ plasmids studied were stable in A. caldus MTH-04. Consequently, this development of a conjugational system for A. caldus MTH-04 will greatly facilitate its genetic study.

Characterization of Antimicrobial Substance Produced by Lactobacillus paraplantarum KNUC25 Isolated from Kimchi (김치로부터 분리된 Lactobacillus paraplantarum KNUC25가 만드는 항균 물질의 특성)

  • Kim, Ma-Rie;Lee, Su-Jin;Seul, Keyung-Jo;Park, Yu-Mi;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.24-32
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    • 2009
  • The KNUC25 strain isolated from over-fermented whole Chinese cabbage kimchi was examined for its physiological characteristics using API 50 CHL system assay and identified as Lactobacillus paraplantarum by analysis of whole-cell protein SDS-PAGE pattern assay and similarity of 16S rDNA sequence. L. paraplantarum KNUC25 had a broad antimicrobial activity spectrum from Gram positive to Gram negative bacteria. Scanning electron micrograph analysis showed that KNUC25 might attack to cell surface of indicator cells and destruction can lead to inhibition of the cell growth. The antimicrobial substance of the KNUC25 strain was stable to various degrading enzymes and at high temperature and not a plasmid-born matter. Resistance to proteolytic enzymes showed that an antimicrobial activity of KNUC25 might not be caused by proteinous substance. Maximum production of antimicrobial substance was the exponential growth phase at $30^{\circ}C$.

Effect of Nifedipine on the Ampicillin Absorption (니페디핀이 암피실린의 흡수에 미치는 영향)

  • Jeong, Hyun-Jeong;Yong, Chul-Soon;Choi, Yoon-Soo;Oh, Doo-Man
    • Journal of Pharmaceutical Investigation
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    • v.27 no.1
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    • pp.57-64
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    • 1997
  • $Amino-{\beta}-lactam$ antibiotics are absorbed by the dipeptide transporter in the small intestine. These uptakes are coupled to a proton influx. The inward proton gradient is partly induced by the $Na^+/H^+$ exchanger and calcium ion is involved in control of this antiport. Interaction between ampicillin which is one of the $Amino-{\beta}-lactam$ antibiotics and nifedipine which is one of calcium channel blocking agents was studied in rats in vivo and with rabbit jejunum mounted on the Sweetana/Grass diffusion cells in vitro. Bioavailability of ampicillin was increased significantly when nifedipine was co-administered orally in rats. There were no differences in the distribution phase and the elimination phase when ampicillin was given either alone or with nifedipine intravenously. Conditions for in vitro experiments were determined. The lift rate of $O_2/CO_2$ gas was controlled to 3 bubbles/sec and ampicillin was stable in the Kreb's buffer at pH 6.0. Absorption of ampicillin was the greatest when the completely-stripped serosal membrane was used. Transport of ampicillin from mucosal to serosal side in the rabbit jejunum was enhanced by 32% in the presence of nifedipine (p=0.059). Above results suggest that nifedipine might increase the plasma level of ampicillin via the improved absorption in the intestine rather than the reduction in the elimination or/and alteration in the distribution.

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Effects of Changes in Glycosylation Sites on Secretion of Recombinant Human Erythropoietin in Cultured CHO Cells

  • Lee, H. G;Lee, P. Y.;Lee, Y. K.;Kim, S. J.;H. K. Chung;M. K. Seo;Park, J. K.;K. S. Min;W. K. Chang
    • Korean Journal of Animal Reproduction
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    • v.27 no.4
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    • pp.299-307
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    • 2003
  • The effects of additions/deletions in glycosylated residues of recombinant human EPO (rhEPO) produced in CHO-K1 on their secretion were examined. hEPO cDNA was amplified from human liver mRNA and cloned into the pCR2.1 TOPO. Using overlapping-extension site-directed mutagenesis method, glycosylation sites at 24th, 38th, 83rd, and 126th were respectively or accumulatively removed by substituting its asparagine (or serine) with glutamine. To add novel glycosylation sites, 69 and 105th leucine was mutated to asparagine. Mutant and wild type rhEPO constructs were cloned into the pcDNA3 expression vector with CMV promoter and transfected into CHO cell line, CHO-K1, to produce mutant rhEPO mutant rhEPO proteins. Enzyme-linked immunosorbant assay (ELISA) and Western analysis with monoclonal anti-EPO antibody were performed using supernatants of the cultures showing transient and stable expressions respectively. Addition of novel glycosylation reduced rhEPO secretion dramatically while deletion mutants had little effect except some double deletion mutants ($\Delta$24/83 and $\Delta$38/83) and triple mutant ($\Delta$24/38/83). This fact suggests that not single but combination of changes in glycosyl groups affect secretion of rhEPO in cell culture, possibly via changes in their conformations.

Improvement of the Negative Plate Performance on Industrial Ni-Zn Battery (산업용 니켈 아연전지 음극성능 향상)

  • Park, Dong-Pil;Kim, Lae-Hyun
    • Journal of Energy Engineering
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    • v.20 no.2
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    • pp.77-83
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    • 2011
  • It is requested to improve negative electrode of Ni-Zn battery for industrial application. Ni-Zn battery has main problems not to commercialize because of short life cycle, heavy gassing and fast electrolyte vaporization so far. It has been studied on 8 cells performances under promoting electric, additional materials and binders changed. With these materials($Ca(OH)_2$, $Bi_2O_3$), negative electrolyte can be manufactured equal and tight as well as low gassing. Furthermore, to supply EG-EP#12(gravity 1.26), keeping stable electrolyte gravity in battery, the life cycle of Ni-Zn battery is extremely improved 200~300% than initial performance.

Nature of a Root-Associated Paenibacillus polymyxa from Field-Grown Winter Barley in Korea

  • RYU CHOONG-MIN;KIM JINWOO;CHOI OKHEE;PARK SOO-YOUNG;PARK SEUNG-HWAN;PARK CHANG-SEUK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.984-991
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    • 2005
  • Soil or seed applications of plant growth-promoting rhizobacteria (PGPR) have been used to enhance growth of several crops as well as to suppress the growth of plant pathogens. In this study, we selected a PGPR strain, Paenibacillus polymyxa strain E681, out of 3,197 heat-stable bacterial isolates from winter wheat and barley roots. Strain E681 inhibited growth of a broad spectrum plant pathogenic fungi in vitro, and treatment of cucumber seed with E681 reduced incidence of damping-off disease caused by Pythium ultimum, Rhizoctonia solani, or Fusarium oxysporum. When inoculated onto seeds as vegetative cells or as endospores, E681 colonized whole cucumber root systems and root tips. Different temperatures such as $20^{\circ}C\;and\;30^{\circ}C$ did not affect root colonization by strain E681. This colonization was associated with a consistent increase in foliar growth of cucumber in the greenhouse. These results indicate that strain E681 is a promising PGPR strain for application to agricultural systems, particularly during the winter season.

A Novel Esterase from Paenibacillus sp. PBS-2 Is a New Member of the ${\beta}$-Lactamase Belonging to the Family VIII Lipases/Esterases

  • Kim, Young-Ok;Park, In-Suk;Nam, Bo-Hye;Kim, Dong-Gyun;Jee, Young-Ju;Lee, Sang-Jun;An, Cheul-Min
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1260-1268
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    • 2014
  • Screening of a gene library from Paenibacillus sp. PBS-2 generated in Escherichia coli led to the identification of a clone with lipolytic activity. Sequence analysis showed an open reading frame encoding a polypeptide of 378 amino acid residues with a predicted molecular mass of 42 kDa. The esterase displayed 69% and 42% identity with the putative ${\beta}$-lactamases from Paenibacillus sp. JDR-2 and Clostridium sp. BNL1100, respectively. The esterase contained a Ser-x-x-Lys motif that is conserved among all ${\beta}$-lactamases found to date. The protein PBS-2 was produced in both soluble and insoluble forms when E. coli cells harboring the gene were cultured at $18^{\circ}C$. The enzyme is a serine protein and was active against p-nitrophenyl esters of $C_2$, $C_4$, $C_8$, and $C_{10}$. The optimum pH and temperature for enzyme activity were pH 9.0 and $30^{\circ}C$, respectively. Relative activity of 55% remained at up to $5^{\circ}C$ with an activation energy of 5.84 kcal/mol, which indicates that the enzyme is cold-adapted. Enzyme activity was inhibited by $Cd^{2+}$, $Cu^{2+}$, and $Hg^{2+}$ ions. As expected for a serine esterase, activity was inhibited by phenylmethylsulfonyl fluoride. The enzyme was remarkably active and stable in the presence of commercial detergents and organic solvents. This cold-adapted esterase has potential as a biocatalyst and detergent additive for use at low temperatures.

Current Status of Gene Therapy as a New Drug Delivery System (신약전달기술체계인 유전자 치료의 현재까지의 개발동향)

  • Bae, Yun-Sung;Cho, Jung-Yoon;Ji, Sang-Mi;Lee, Young-Joo
    • Journal of Pharmaceutical Investigation
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    • v.32 no.3
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    • pp.153-159
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    • 2002
  • Gene therapy is fundamentally a sophisticated drug delivery technology to cure a disease by the transfer of genetic material to modify living cells. In other words, the gene is used as a therapeutic drug much like a chemical compound is employed in chemotherapy. Currently almost 600 clinical trials are underway worldwide since the first clinical trials carried out in 1990 to treat adenosine deaminase deficiency using retroviral vectors. Despite the great progress still is there no gene therapy product being approved as a new drug. This is partly due to a lack of an ideal gene delivery system that is safe and can provide stable, optimal level production of the therapeutic proteins in the cell. This review covers the current status of several different biological and physico-chemical agents that are being developed as gene delivery vehicles. Although gene therapy promises great hopes toward the cure of a broad spectrum of genetic and acquired diseases, the success of gene therapy heavily asks for the development of vector systems for safe and efficient application in humans.

Configuration and Ground Tests of Solar Cell and Fuel Cell Powered System for Long Endurance UAV (장기체공 무인기용 태양전지-연료전지를 활용한 동력원 구성 및 지상시험)

  • Park, Byeongseob;Kim, Hyuntak;Baek, Seungkwan;Kwon, Sejin
    • Journal of the Korean Society of Propulsion Engineers
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    • v.19 no.4
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    • pp.94-101
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    • 2015
  • Each of power systems of solar cell and fuel cell were configured and validated for long endurance UAV, as the preliminary research for the integration of power systems. Solar power system consisted of solar modules fabricated by solar cells of Sunpower's C60, commercial solar MPPT controller and Li-po battery, and then was validated. The re-start characteristics of hydrogen production from $NaBH_4$ hydrolysis was validated for operating the commercial fuel cell. The average voltage drop of Li-po battery in solar power system was -2.9 V/hour. The performance of re-start characteristics of $NaBH_4$ hydrolysis was stable in sequence mode of mission profile. Each of single systems were satisfied for the proposed mission profile.

Haematologic Parameters in Metastatic Colorectal Cancer Patients Treated with Capecitabine Combination Therapy

  • Inanc, Mevlude;Duran, Ayse Ocak;Karaca, Halit;Berk, Veli;Bozkurt, Oktay;Ozaslan, Ersin;Ozkan, Metin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.253-256
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    • 2014
  • Background: The standard treatment in the metastatic colorectal cancer consists of 5-FU based infusional regimens. However, with oral fluoropyrimidines, equal tumor responses may be obtained. Capecitabine causes macrocytosis of the cells by inhibition of DNA synthesis. In this context, a relationship was found between mean corpuscular volume (MCV) and response to therapy in breast cancer patients treated with Capecitabine, but whether this relationship also pertains in colorectal cancer has not been established. Materials and Methods: A total of 102 metastatic colorectal cancer patients treated with a oxaliplatin (XELOX)${\pm}$Bevacizumab combination were retrospectively evaluated. Patients were randomized into three groups. Hematological parameters (MCV, MPV, PCT, PLT, NLR) were recorded retrospectively, before treatment and after 3 cycles of chemotherapy. Results: After three cycles of therapy, 20 (19.6%) patients had progressive disease (PD), 41 (40.1%) had stable disease (SD), and 41 (40.1%) demonstrated a partial response (PR). In 62 (60.7%) treatment was with capesitabin plus XELOX therapy, and in 40 (39.2%) it was XELOX-Bevacizumab combination therapy. There was no difference among three groups before the treatment in terms of MCV, MPV, PCT, PLT, and NLR. MCV showed significant increase in chemotherapy response groups (PR and SD). In addition, a significant decrease was observed for platelet count in chemotherapy response groups. While NLR decrease was seen in only a PR group, PCT decrease was observed in all three groups. PCT and PLT values were higher in patients receiving Bevacizumab. Conclusions: PLT, PCT, MPV, and NLR values were decreased due to Capecitabine-based chemotherapy, however MCV was increased. PCT and PLT values were higher in patients who received Bevacizumab than those who did not. MCV, PLT, and NLR can be considered as important factors in predicting response to colorectal carcinoma treatment.