• Title/Summary/Keyword: negative mutants

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Variations in the Chemical Compositions of Peanut Mutants Induced by Gamma Radiation

  • Doo, Hong Soo;Cheong, Young Keun;Paik, Ki Hun
    • Korean Journal of Breeding Science
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    • v.40 no.2
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    • pp.113-118
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    • 2008
  • This study was carried out to investigate the chemical composition of peanut mutants induced by gamma radiation (Co-60) at 300 Gy in seeds of the cultivar Shinnamkwang. The protein contents of twenty-eight peanut mutants ranged from 23.3% to 31.7% and were increased by from 0.5% to 8.4% in fifteen mutants lines from the 26.8% of the original variety, cv. Shinnamkwang. Lipid contents of in mutants ranged from 43.2% to 53.5%, an increase of 0.2% to 5.7% from the 47.8% of the original variety. The range of unsaturated oleic acid in 28 mutants was from 38.9% to 56.9% an increase of from 1.3% to 14.0% from the 50.6% in the original variety. Linoleic acid, the highest unsaturated fatty acid, constituted 32.6% meanly of mutants, it was 17.4% lesser than oleic acid, ranging from 25.9% to 42.0%. Palmitic acid (16:0) contents ranged from 8.6% to 11.1%, and the mutant line-9 had the highest content. The ratio of oleic to linoleic acid was 0.9 to 2.2. A negative coefficient (r=-0.98**) was obtained between oleic and linoleic acid, but for other fatty acids, no significant relation was observed. Similarly, a negative coefficient of r=-0.68**was observed between saturated and unsaturated acids. The sucrose compositions of mutants ranged from 2.6% to 6.2%.

Effect of Gluconic Acid on the Production of Cellulose in Acetobacter xylinum BRC5

  • PARK, SANG TAE;TAEKSUN SONG;YOUNG MIN KIM
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.683-686
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    • 1999
  • Four mutants of Acetobacter xylinum BRC5 defective in gluconic acid production were isolated from UV-irradiated cells. The gluconic acid-negative mutants did not show glucose oxidase activity. The mutants were also defective in cellulose production. A randomly selected mutant grown in the Hestrin-Schramm medium (pH 6.0) supplemented with gluconic acid, however, was found to synthesize cellulose. The mutant grown in Hestrin-Schramm medium whose pH was adjusted to 5.0 with HC1 and contained no gluconic acid also produced cellulose. Wild-type cells grown under the same condition synthesized cellulose more rapidly than those grown in the pH 6.0 medium.

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Functional Abnormalities of HERG Mutations in Long QT Syndrome 2 (LQT2)

  • Hiraoka, Masayasu
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.5
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    • pp.367-371
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    • 2001
  • The chromosome 7-linked long QT syndrome (LQT2) is caused by mutations in the human ether-a- go-go-related gene (HERG) that encodes the rapidly activating delayed rectifier $K^+$ current, $I_{Kr},$ in cardiac myocytes. Different types of mutations have been identified in various locations of HERG channel. One of the mechanisms for the loss of normal channel function is due to membrane trafficking of channel protein. The decreased channel function in some deletion mutants appears to be due to loss of coupling with wild type HERG to form the functional channel as the tetramer. Most of missense mutants with few exceptions could interact with wild type HERG to form functional tetramer and caused dominant negative suppression with co-injection with wild type HERG showing variable effects on current amplitude, voltage dependence, and kinetics of activation and inactivation. Two missense mutants at pore regions of HERG found in Japanese LQT2 (A614V and V630L) showed accentuated inward rectification due to a negative shift in steady-state inactivation and fast inactivation. One mutation in S4 region (R534C) produced a negative shift in current activation, indicating the S4 serving as the voltage sensor and accelerated deactivation. The C-terminus mutation, S818L, could not express the current by mutant alone and did not show dominant negative suppression with co-injection of equal amount of wild type cRNA. Co-injection of excess amount of mutant with wild type produced dominant negative suppression with a shift in voltage dependent activation. Therefore, multiple mechanisms are involved in different mutations and functional abnormality in LQT2. Further characterization with the interactions between various mutants in HERG and the regulatory subunits of the channels (MiRP1 and minK) is to be clarified.

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The effect of surface charge balance on thermodynamic stability and kinetics of refolding of firefly luciferase

  • Khalifeh, Khosrow;Ranjbar, Bijan;Alipour, Bagher Said;Hosseinkhani, Saman
    • BMB Reports
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    • v.44 no.2
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    • pp.102-106
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    • 2011
  • Thermodynamic stability and refolding kinetics of firefly luciferase and three representative mutants with depletion of negative charge on a flexible loop via substitution of Glu by Arg (ER mutant) or Lys (EK mutant) as well as insertion of another Arg in ER mutants (ERR mutant) was investigated. According to thermodynamic studies, structural stability of ERR and ER mutants are enhanced compared to WT protein, whereas, these mutants become prone to aggregation at higher temperatures. Accordingly, it was concluded that enhanced structural stability of mutants depends on more compactness of folded state, whereas aggregation at higher temperatures in mutants is due to weakening of intermolecular repulsive electrostatic interactions and increase of intermolecular hydrophobic interactions. Kinetic results indicate that early events of protein folding are accelerated in mutants.

Effects of Mixing Conditions on the Production of Microbial Cellulose by Acetobacter xylinum

  • Lee, Hei-Chan;Xia Zhao
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.4 no.1
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    • pp.41-45
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    • 1999
  • Microbial cellulose has many potential applications due to its excellent physical properties. The production of cellulose from Acetobacter xylinum in submerged culture is, however, beset with numerous problems. The most difficult one has been the appearance of negative mutants under shaking culture conditions, which is deficient of cellulose producing ability. Thus genetic instability of Acetobacter xylinum under shaking culture condition made developing a stable mutant major research interest in recent years. To find a proper type of bioreactor for the production of microbial cellulose, several production systems were developed. Using a reactor system with planar type impeller with bottoms sparging system, it was possible to produce 5 g/L microbial cellulose without generating cellulose minus mutants, which is comparable to that of static culture system.

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Thermosensing of Thermotactic Mutants, Dictyostelium discoideum Amoebae in Vegetative Stage

  • Hong, Choo-Bong
    • Journal of Plant Biology
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    • v.26 no.3
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    • pp.131-139
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    • 1983
  • Temperature response of amoebae of thermotactic mutants have been investigated. Amoebae of the mutant strain HO 428 showed positive thermotaxs which is strong at lower temperaturs and drops sharply above the growth temperature of amoebae. The temperature response of HO 428 amoebae was not affected by the length of amoebae on the grdients. HO 596 amoebae seemed to have both positive and negative thermotactic responses shortly after food depletion. Longer exposure of these amoebae on the thermal gradients induced a stronger negative response at lower temperatures and an apparent positive response at higher temperatures. A similar changes could be observed in HO 1445 amoebae. Based on the steady positive thermotactic response by HO 428 amoebae and the mode of change in termperature response at higher temperatures, 24$^{\circ}C$ and $25^{\circ}C$, by HO 596 amoebae, a model for the temperature response of vegetative Dictyostelium discoideum amoebae, strain HL 50, has been proposed. The main features of the model are: a positive response at the thermal gradients with midpoint temperatures lower than the growth temperatures of amoebae and a negative response above it.

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Improvement of Permeability to Organic Solvent in Escherichia coli for a Toxicity Biosensor

  • Bae, Hee-Kyung;Shin, Pyong-Kyun;Song, Bang-Ho
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2001.06b
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    • pp.14-16
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    • 2001
  • The outer membrane (OM) of gram-negative bacteria acts as an effective permeability barrier against noxious agents including several antibiotics and organic solvents, and lipopolysaccharide (LPS) is the key molecule for this function. Outer membrane modified mutants (Ml-166, M2-42, M3-21) of E. coli DH5$\alpha$/pBSl were selected through a mutation using EMS (ethyl-methane-sulfonate). Among the selected mutants, M3-21 was twice as sensitive as LumisTo $x^{ }$ to benzene and M2-41 was 8 times as sensitive as LumisTo $x^{ }$ to toluene. To identify the structural change in the membrane by mutation, the relative cell surface hydrophobicities and the absorption of the crystal violet to the organisms were measured. All the mutants absorbed more crystal violet than their parent and the absorption of crystal violet increased in cell walls as carbohydrate of lipopolysaccharide decreased. When the cell surface hydrophobicities of DH5/pBSl and its mutants were measured by the BATH, the hydrophobicities of mutants increased compared to their parent in several organic solvents. The difference of lipopolysaccharide between DH5/pBSl and its mutants was identified by various ways such as the SDS-PAGE gel, the screening of LPS molecular weights, the mass spectrometry, and MALDI-TOF.F.

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Conversion of G. hansenii PJK into Non-cellulose-producing Mutants According to the Culture Condition

  • Park, Joong-Kon;Hyun, Seung-Hun;Jung, Jae-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.5
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    • pp.383-388
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    • 2004
  • The conversion of a cellulose-producing cell ($Cel^+$) from Gluconacetobacter hansenii PJK (KCTC 10505 BP) to a non-cellulose-producing cell ($Cel^-$) was investigated by measuring the colony forming unit (CFU). This was achieved in a shaking flask with three slanted baffles, which exerted a strong shear stress. The addition of organic acid, such as glutamic acid and acetic acid, induced the conversion of microbial cells from a wild type to $Cel^-$ mutants in a flask culture. The supplementation of $1\%$ ethanol to the medium containing an organic acid depressed the con-version of the microbial cells to $Cel^-$ mutants in a conventional flask without slanted baffles. The addition of ethanol to the medium containing an organic acid; however, accelerated the conversion of microbial cells in the flask with slanted baffles. The $Cel^+$ cells from the agitated culture were not easily converted into $Cel^-$ mutants on the additions of organic acid and ethanol to a flask without Slanted baffles, but some portion of the $Cel^+$ cells were converted to $Cel^-$ mutants in a flask with slanted baffles. The conversion ratio of $Cel^+$ cells to $Cel^-$ mutants was strongly re-lated to the production of bacterial cellulose independently from the cell growth.

Effects of Natural Selection, Mutagenesis, and Protoplast Formation and Cell Wall Regeneration on the Production of Aminoglycoside Antibiotics

  • Goo, Yang-Mo;Lim, Hyon-Joo;Lim, Seok-Ran;Kim, Kong-Hwan;Lim, Bun-Sam;Lee, Sae-Bae
    • Archives of Pharmacal Research
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    • v.12 no.4
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    • pp.249-253
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    • 1989
  • High producers or blocked mutants of aminoglycoside antibiotic-producing Streptomyces spp. were selected by application of an agar plug method and by culturing individual colonies in broth. The productivities of aminoglycoside antibiotic producing organisms were increased by selection of a high producer from colonies obtained by spreading spores of wild strain, or survived from treatment of a mutagen or from the colonies regenerated from protoplast-formation and cell-wall regenerations. Some mutagen treated colonies lost the ability to produce antibiotics (5-8%). Some A-factor negative and deostreptamine or streptidine negative mutants were obtained by N-methyl-N'-nitro-N-nitrosomethylguanidine (MNNG) treatment. Many of the survivors from the MNNG treatment lost the ability to produce antibiotics. Major colonies produced less amount of antibiotics ; only few survived colonies produced more antibiotics than the parent. Resistance of Streptomyces spp. against the antibiotics produced by itself was also markedly affected by mutagen treatment.

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Putative Negative Regulation of Novel MarB along with MarA upon the Function of AcrAB/TolC Efflux Pump of Escherichia coli K-12 (대장균 K-12의 AcrAB/TolC Efflux Pump의 기능에 대한 MarB와 MarA의 추정적 억제조절)

  • Byung-Tae Park
    • Biomedical Science Letters
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    • v.5 no.1
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    • pp.27-40
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    • 1999
  • This study was focused on the evaluation of MarB alongside with MarA for its regulatory effects upon the efflux function of AcrAB pump, which were induced or not, perhaps as a target. Transductions of marR and/or acrAB mutation which were derived from Mar and/or AcrAB mutants of wild type E. coli K-12, respectively, into the multicopy plasmid in wild type E. coli backgrounds or into the chromosome of isogenic parents were done. Minimal inhibitory concentration (MIC) of transduced mutants was compared with their original mutants. This study reports the indirect evidences that suggests a model in which MarB along with MarA have a putative negative regulatory effect upon the efflux function of AcrAB/TolC pump while MarA alone have a positive regulatory effect to the expression of acrRAB operon at transcription level. The target of MarB with MarA for its putative negative regulator might be the AcrAB efflux pump. Another efflux system (s) might be negatively regulated by MarB with MarA, and be involved in the efflux of antibiotics which were otherwise extruded preferentially by AcrAB efflux pump.

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