• Title/Summary/Keyword: New mutant

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Action Mechanism of LB10522, a New Catechol-Substituted Cephalosporin (카테콜 치환체를 가진 세파로스포린계 항생제 LB10522의 작용기전)

  • Kim, Mu-Yong;Oh, Jeong-In;Paek, Kyoung-Sook;Kim, In-Chull;Kwak, Jin-Hwan
    • YAKHAK HOEJI
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    • v.40 no.1
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    • pp.102-111
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    • 1996
  • LB10522 is a new parenteral broad spectrum cephalosporin with a catechol moiety at C-7 position of beta-lactam ring. This compound can utilize tonB-dependent iron transp ort system in addition to porin proteins to enter bacterial periplasmic space and access to penicillin-binding proteins (PBPs) which are the lethal targets of ${\beta}$-lactam antibiotics. The chelating activity of LB10522 to metal iron was measured by spectrophotometrically scanning the absorbance from 200 to 900nm. When $FeCl_3$ was added, optical density was increased between 450 and 800nm. LB10522 was more active against gram-negative strains in iron-depleted media than in iron-replete media. This is due to the increased expression of iron transport channels in iron-depleted condition. LB10522 showed a similar activity against E. coli DC2 (permeability mutant) and E. coli DCO (wild type strain) in both iron-depleted and iron-replete media, indicating a minimal permeaility barrier for LB10522 uptake. LB10522 had high affinities to PBP 3 and PBP 1A, 1B of E. coli. By blocking these proteins, LB10522 caused inhibition of cell division and the eventual death of cells. This result was correlated well with the morphological changes in E. coli exposed to LB10522. Although the in vitro MIC of LB10522 against P. aeruginosa 1912E mutant (tonB) was 8-times higher than that of the P. aeruginosa 1912E parent strain, LB10522 showed a similar in vivo protection efficacy against both strains in the mouse systemic infection model. This result suggested that tonB mutant, which requires a high level of iron for normal growth, might have a difficulty in surviving in their host with an iron-limited environment.

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Induction of Chlorophyll Deficient Mutant Plant of Cymbidium kanran by EMS Treatment (EMS처리에 의한 한란의 엽록소 결핍 돌연변이 식물체의 유도)

  • 이효연;정재성;이종석
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.183-187
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    • 1998
  • Chlorophyll mutants were produced by treating the rhizome of Cymbidium kanran with mutagen, EMS(ethyl methan sulfonate). The germination ratio of Cymbidium kanran seeds was 5.5 times higher when the seeds were treated with ultrasonic treatment for 20 minutes than untreated control. Fifty to sixty percent of the rhizomes became dark brown when they were cultured in a liquid growth medium containing 0.2% EMS for three weeks. When the dark-brown rhizomes were cultured in a solified MS medium, new rhizomes were developed from a part of the old ones. Chlorophyll mutant rhizomes were obtained from a meristem tissue by a subculturing the cuts of these new rhizomes for a year. Of the chlorophyll mutants, a zigzag-striped type of rhizome was dominant and light-yellow and albino ones were also produced. While the zigzag-striped type rhizomes were differentiated into green and striped plant, the light yellow and the white rhizomes produced yellow-striped and albino plants repectively.These results indicate that the EMS treatment on the rhizome is an effective means to induce a chlorophyll mutant. We believe that this method may be useful to produce variegated plants chlorophyll mutants from other orchids.

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Studies on the Mutation Breeding of Naked Barley I. A New Mutant Variety "Radiation No. 6" and Several Promising Mutant Lines by Radiation (쌀보리의 돌연변이육종에 관한 연구 I. 방사선에 의한 유용변이계통 및 신품종 ′방사6호′에 대하여)

  • Kim, Y.S.;Park, K.Y.;Lee, D.K.;Kim, I.H.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.21 no.1
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    • pp.82-86
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    • 1976
  • 'Radiation No. 6' the first mutant variety of naked barley in Korea was developed from Bangju as .a mother variety. The new variety was characterized by early maturity, short-culm and stablized high yield by the increased number of spikes. 'Radiation No. 6' was registered as a leading variety in 1974, .after 7 years since induction of mutation started.of mutation started.

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A New Function of Skp1 in the Mitotic Exit of Budding Yeast Saccharomyces cerevisiae

  • Kim, Na-Mil;Yoon, Ha-Young;Lee, Eun-Hwa;Song, Ki-Won
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.641-648
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    • 2006
  • We previously reported that Skp1, a component of the Skp1-Cullin-F-box protein (SCF) complex essential for the timely degradation of cell cycle proteins by ubiquitination, physically interacts with Bfa1, which is a key negative regulator of the mitotic exit network (MEN) in response to diverse checkpoint-activating stresses in budding yeast. In this study, we initially investigated whether the interaction of Skp1 and Bfa1 is involved in the regulation of the Bfa1 protein level during the cell cycle, especially by mediating its degradation. However, the profile of the Bfa1 protein did not change during the cell cycle in skp1-11, which is a SKP1 mutant allele in which the function of Skp1 as a part of SCF is completely impaired, thus indicating that Skp1 does not affect the degradation of Bfa1. On the other hand, we found that the skp1-12 mutant allele, previously reported to block G2-M transition, showed defects in mitotic exit and cytokinesis. The skp1-12 mutant allele also revealed a specific genetic interaction with ${\Delta}bfa1$. Bfa1 interacted with Skp1 via its 184 C-terminal residues (Bfa1-D8) that are responsible for its function in mitotic exit. In addition, the interaction between Bfa1 and the Skp1-12 mutant protein was stronger than that of Bfa1 and the wild type Skp1. We suggest a novel function of Skp1 in mitotic exit and cytokinesis, independent of its function as a part of the SCF complex. The interaction of Skp1 and Bfa1 may contribute to the function of Skp1 in the mitotic exit.

Polyvinyl Alcohol 분해 공생 균주에 의한 염색 폐수 중의 PVA 제거

  • Kim, Chul Ki;Choi, Yong-Jin;Lee, Chul-Woo;Rim, Yeon-Taek;Ryu, Jae-Keun
    • Microbiology and Biotechnology Letters
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    • v.25 no.1
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    • pp.89-95
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    • 1997
  • The current processer of the textile wastewater treatment are mostly consisted of a combination of a physico-chemical and a biological treatment. The overall efficiency of these processes is, however, assessed to be fairly low. It is even worse during the summer season when temperature of the wastewater rises above 40$\circ $C. Therefore, a feasible process of the textile wastewater treatment which can work efficiently at higher temperatures was investigated in this work. We used a bench scale reactor consisted of one 4 liter anaerobic and one 8 liter aerobic tank, and the thermophilic symbiotic PVA degraders, Pasteruella hemolytica KMG1 and Pseudomonas sp. KMG6 that had been isolated in our laboratory. In the preliminary flask experiments, we observed that the thermophilic symbiotic PVA degraders could not grow in the wastewater substrate. Then, we isolated the mutant strains by acclimating the KMG1and KMG6 strains to the wastewater medium. The mutant symbionts (KMG1-1 and KMG6-1) were isolated through 6 times successive transfers into the fresh wastewater medium after 5 days culture for each. The mutant strains obtained grew well in the mixed medium composed of 75% wastewater and 25% synthetic medium, and supplemented with 0.5% PVA as a sole carbon source. During the culture for 14 days at pH 7.0 and 40$\CIRC $C, the bacteria assimilated about 89% of the added PVA. The symbionts degraded equally well all the PVA substrates of different molecular weight (nd=500~30000). In contrast to the flask experiments, in the reactor system the mutant strains showed very low levels of the PVA and COD removal rates. However, the new reactor system with an additional aerobic tank attained 82% removal rate of COD, 94% of PVA degradation and 71% of color index under the conditions of 5% inoculm on the tank 2, incubation temperature of 40$\circ $C, dissolved oxygen level of 2~3 mg/l and retention time of 30 hours. This result ensures that the process described above could be an efficient and feasible treatment for the PVA contained textile wastewater at higher temperatures.

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Auxin Induced Expression of Expansin is Alered in a New Aux1 Allele that Shows Severe Defect in Gravitropic Response

  • Jeong, Hae-Jun;Kwon, Ye-Rim;Oh, Jee-Eun;Kim, Ki-Deok;Lee, Sung-Joon;Hong, Suk-Whan;Lee, Ho-Joung
    • Journal of Applied Biological Chemistry
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    • v.49 no.4
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    • pp.148-153
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    • 2006
  • While the underlying molecular mechanism remains to be elucidated, recent studies suggest that polar auxin transport is a key controlling factor in triggering differential growth responses to gravity. Identification of regulatory components in auxin-mediated differential cell expansion would improve our understanding of the gravitropic response. In this study, we identify a mutant designated aux1-like(later changed to aux1), an allele of the aux1 mutant that exhibits a severely disrupted root gravitropic response, but no defects in developmental processes. In Arabidopsis, AUX1 encodes an auxin influx carrier. Since in-depth characterization of the gravitropic response caused by mutations in this gene has been performed previously, we focused on identifying the downstream genes that were differentially expressed compared to wild-type plants. Consistent with the mutant phenotype, the transcription of the auxin-responsive genes IAA17 and GH3 were altered in aux1 plants treated with IAA, 2, 4-D and NAA. In addition, we identified two expansin genes EXP10 and EXPL3 that exhibited different expression in wild-type and mutant plants.

New Classification of Plasmodiophora brassicae Races Using Differential Genotypes of Chinese Cabbage

  • Kim, Hun;Choi, Gyung Ja
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.28-28
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    • 2015
  • Clubroot disease caused by Plasmodiophora brassicae induces severe losses of cruciferous vegetables worldwide. To control clubroot of Chinese cabbage, many CR (clubroot resistance) F1 hybrid cultivars have been bred and released in Korea, China and Japan. In this study, we determined the race of P. brassicae 12 field isolates, which collected from 10 regions in Korea, using Williams' differential varieties including two cabbage ('Jersey Queen', 'Badger Shipper') and two rutabaga ('Laurentian', 'Whilhelmsburger'). By Williams' differential varieties, 12 clubroot pathogens were assigned into one (GN2), two (HS and YC), two (HN1 and HN2), three (DJ, KS and SS) and four (GS, GN1, JS and PC) isolates for races 1, 2, 4, 5 and 9, respectively. In addition, the degree of resistance of 45 CR cultivars that were from Korea, China and Japan was tested with the 12 isolates. The 45 CR cultivars of Chinese cabbage were differentiated into three genotypes according to their resistance responses. Even though the 12 P. brassicae isolates were same race by Williams' differential varieties, three CR genotypes showed different resistance response to the isolates. These results indicate that races of P. brassicae by Williams' differentials were not related with resistance of CR cultivars, and three CR genotypes represented qualitative resistance to the P. brassicae isolates. CR genotype I including 'CR-Cheongrok' showed resistance to GN1, GN2, JS, GS, HS, DJ and KS isolates and susceptibility to YC, PC, HN1, HN2 and SS isolates. And CR genotype II such as 'Hangkunjongbyungdaebaekchae' was resistant to GN1, GN2, JS, GS, HS, YC, PC and HN1 and susceptible to DJ, KS, SS and HN2. CR genotype III including 'Chunhajangkun' and 'Akimeki' represented resistance to 10 isolates except for SS and HN2 isolates. Based on these results, we selected 'CR-Cheongrok', 'Hangkunjongbyungdaebaekchae', and 'Chunhajangkun' as a representative cultivar of three CR genotypes and 'Norangkimjang' as a susceptible cultivar. Furthermore, we investigated the resistance of 15 lines of Chinese cabbage, which were provided by seed companies, to 11 isolates except for HN1 of P. brassicae. The results showed that three lines were susceptible to all the tested isolates, whereas five, four, and three lines represented the similar responses corresponding to the CR genotypes I, II, and III, respectively; there is no line of Chinese cabbage showing different resistance patterns compared to three CR genotypes. In particular, line 'SS001' showing resistance responses of CR genotype II was a parent of 'Saerona' that have been commercialized as a CR $F_1$ cultivar of Chinese cabbage. Together, we divided 12 isolates of P. brassicae into 4 races, designated by wild type, mutant type 1, mutant type 2, and mutant type 3. Wild type including GN1, GN2, JS, GS, and HS isolates of P. brassicae was not able to infect all the cultivars of three CR genotypes, whereas, mutant type 3 such as SS and HN2 isolates developed severe clubroot disease on all the CR genotype cultivars. To mutant type 1 including DJ and KS isolates, CR genotypes I, II and III were resistant, susceptible and resistant, respectively. In contrast, to mutant type 2 including YC, PS, and HN1 isolates, CR genotypes I, II and III showed susceptibility, resistance and resistance, respectively. Taken together, our results provide the extended knowledge of classification of P. brassicae races, which is useful information for the breeding of resistant crops, with a suggestion that 'Norangkimjang', 'CR-Cheongrok', 'Saerona' and 'Chunhajangkun' cultivars of Chinese cabbage could be used as new race differentials of P. brassicae for clubroot disease assay.

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Induction of apoptosis by etoposide treatment in colon cancer cell line SNU C2A (대장암 세포주 SNU C2A에서 etoposide 처리에 의한 apoptosis 유도)

  • Jung, Ji-Yeon;Na, Yun-sook;Jung, Ho-Chul;Oh, Sang-Jin
    • IMMUNE NETWORK
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    • v.1 no.3
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    • pp.221-229
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    • 2001
  • Background: Inactivation of tumor suppressor genes is believed to be important in the development of many human malignancies. Recently, several lines of evidence have indicated that the wild type p53 gene located at 17p13.3, may function as a tumor suppressor gene and that a mutant p53 gene could promote transformation by inactivating normal p53 function in a dominant negative fashion. These broad spectrum of p53 mutation in human cancers provide that mutant p53 and their protein may be potential targets of tumor diagnostic and therapeutic interventions. Method: Colony formation was performed to investigate growth suppressional ability. p53 expression pattern was examined by western blot and p53-mediated transactivation ability was assessed by CAT activity. SNU C2A cells were observed in apoptotic aspects induced by etoposide and $H_2O_2$ treatment, detecting sensitivity on agent, DNA fragmentation through agarose gel, chromatin condensation by fluorescence microscope, and cell cycle distribution by FACS. Result: 1) p53 mutant his179arg ($histidine{\rightarrow}arginine$) detected in SNU C2A cells lost transcriptional activity and growth suppression ability, showing dominant negative effect on its wild type p53. 2) Etoposide-treated SNU C2A cells induced apoptosis, exhibiting dramatic reduction of cell growth, DNA fragmentation, nuclear condensation formation of apoptotic body and increment of sub-G1 cell fraction. 3) Etoposide and $H_2O_2$-treated SNU C2A cells have no high increase of p53 expression and overexpressed p53 protein changed localization, from cytoplasm to nucleus. Also, p53-mediated transcriptional activity was increased by agents-treatment. Conclusion: SNU C2A cells coexpress wild-type and mutant p53 protein induced apoptosis in the condition on DNA damage, through localizational shift from cytoplasm to nucleus of p53 protein rather than the induction of p53 protein. SNU C2A cells derived mutant p53 his179arg abrogated both the growth supression ability and transactivational activity, showing inhibition effect on transcriptional activity of wild type p53, but did not repress the activity of wild type p53 in SNU C2A cells owing to dominant activity of wild type. These cell condition may provide new gene therapeutic implications leading effective antiproliferation of cell when mutant and wild-type p53 protein were co-expressed in cell.

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Control of Ragweed (Ambrosia artemisiifolia) of Mutant N-29 Broth Filtrate of Streptomyces scopuliridis KR-001 (토양 방선균 N-29 배양 여액의 생태계교란 식물 돼지풀 방제효과)

  • Kim, Jae-Deok;Kim, Young-Sook;Kwak, Hwa-Sook;Kim, Hye-Jin;Lee, Youn-Me;Ko, Young-Kwan;Park, Kee-Woong;Choi, Jung-Sup
    • Weed & Turfgrass Science
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    • v.7 no.3
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    • pp.219-229
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    • 2018
  • We isolated Streptomyces scopuliridis KR-001 that produced herbicidal active metabolite(s) against several grass and broad leaf weeds. In this study, potential as natural herbicide of mutant N-29 broth filtrate of S. scopuliridis KR-001 was investigated to Ambrosia artemisiifilia in a greenhouse and field condition. The broth filtrate of mutant N-29 by foliar application showed a strong herbicidal activity to A. artemisiifilia with leaf stage in a greenhouse condition. Also, field trial of foliar application within treatment range had effectively controlled with early and middle stage of A. artemisiifilia at the natural habitats. Phytotoxic symptoms of mutant N-29 broth filtrate by foliar application were wilting and discoloration, and burn-down of leaves and finally plant death. These results suggest that mutant N-29 broth filtrate is considered possible as a natural herbicide for controlling environmentally friend to invasive alien plant such as A. artemisiifilia and may provide a new lead molecule for a more efficient herbicide.

Isolation of a Mutant with Thermotolerance and Ethanol Tolerance Using Proofreading-deficient DNA Polymerases in Saccharomyces cerevisiae (출아효모에서 proofreading-deficient DNA polymerase를 이용한 내열성 및 에탄올내성 변이 주의 분리)

  • Kim, Yeon-Hee
    • Journal of Life Science
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    • v.29 no.8
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    • pp.916-921
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    • 2019
  • In this study, we constructed a biological system that exhibited thermotolerance, ethanol tolerance, and increased ethanol productivity using a random mutagenesis method. We attempted to isolate a thermotolerant mutant using proofreading-deficient DNA polymerase ${\delta}$ and ${\varepsilon}$ encoded by the pol3 and pol2 genes, respectively, in Saccharomyces cerevisiae. To obtain mutants that could grow at high temperatures ($38^{\circ}C$ and $40^{\circ}C$), random mutagenesis of AMY410 (pol2-4) and AMY126 (pol3-01) strains was induced. The parental strains (AMY410 and AMY126) grew poorly at temperatures higher than $38^{\circ}C$. By stepwise elevation of the incubation temperature, AMY410-Ht (heat tolerance) and AMY126-Ht strains that proliferated at $40^{\circ}C$ were obtained. These strains were further incubated in medium containing 6% and 8% ethanol and then AMY410-HEt (heat and ethanol tolerance) and AMY126-HEt strain with ethanol tolerance at an 8% ethanol concentration was obtained. The AMY126-HEt strain grew even at an ethanol concentration of 10%. Furthermore, following the addition of high concentrations of glucose (5% and 10%), an AMY126-HEt3 strain with increased ethanol productivity was isolated. This strain produced 24.7 g/l of ethanol (95% theoretical conversion yield) from 50 g/l of glucose. The findings demonstrate that a new biological system (yeast strain) showing various phenotypes can be easily and efficiently bred by random mutagenesis of a proofreading- deficient mutant.