• 제목/요약/키워드: ts mutants

검색결과 19건 처리시간 0.019초

Bacillus thringiensis var. kurstaki 감온성 돌연변이주의 일부특성 (Partial Characterization of Bacillus thuringiensis var. kurstaki Temperature-sensitive Mutants)

  • 김영권;유관희;이형환;이호원
    • 미생물학회지
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    • 제23권3호
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    • pp.203-208
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    • 1985
  • Bacillu! thuringiensis var. krustaki 3ab k-3의 감온성치사돌연변이체에 대한 생물학적 성상의 변이성의 차이를 조사하고 유전적 특성을 규명하기 위하여 생화학적 성상검사, 항생제에 대한 감수성 조사, 용혈성 검사, 돌연변이체균주의 특수배지에서의 성장비교, Mouse에 대한 치사효과를 $28^{\circ}C$의 허용온도에서 조사하였다. Urease를 생산하지 못하는 돌연변이체는 ts-U154, ts-U601, ts-U602, ts-U603, ts-U604, t-U788 이었다. ts-U603만이 운동성이 상실되었고, ts-U154는 Salicin과 cellobiose을 분해하지 못했고, ribose를 분해하지 않는 균주는 ts-U603였다. 항생제에 대한 반응은 ampicillin, cephalothin, penicillin에는 모두 저항성을 나타냈으나, ts-U154 균주만이 감수성을 나타냈다. Fructus gardenia배지에서 색소를 생산하지 않은 것은 ts-U21, ts-U74, ts-U131과 ts-U154였다 가장 적은 용혈성을 나타내는 균주는 정상형고 ts-U602였고 가장 많은 용혈성을 나타내는 것은 ts-U603 (1:512 x) 이었다. Mouse 에 대한 치사효과는 전혀 없었다.

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Bacillus sphaericus의 치사감온성 돌연변이체의 분리 (Isolation of Conditional Lethal Temperature-sensitive Mutants of Bacillus sphaericus)

  • 김영한;이형환
    • 한국미생물·생명공학회지
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    • 제13권1호
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    • pp.41-49
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    • 1985
  • Bacillus sphaericss 1593균주를 ultra violet light (366nm, 253.7nm)와 dimethyl sulfate으로 처리하여 다수의 감온성 돌연변이체를 분리하여 이중 25균주를 선택하였고 24개의 감온성 돌연변이체는 $42^{\circ}C$의 제한온도에서 전혀 성장을 하지 않았으며, 9개의 감온성 돌연변이체는 제한온도 $42^{\circ}C$에서 허용온도의 성장과 비교하여 1/2정도의 감소를 보였다. 기타 2개의 감온성 돌연변이체는 저온 민감성인 감온성 돌연변이체로서 허용온도($30^{\circ}C$)에서의 성장이 제한온도($42^{\circ}C$) 성장보다 감소를 보였다. 이러한 결과에 따라서 감온성 돌연변이체를 치사군과 비치사군 및 저온민감성 감온성 돌연변이체로 분리할 수 있었다. 또한 치사군의 제한온도에서의 발육은 고체배지 및 액체배지 양쪽에서 성장을 하지 않는 반면 비치사군들의 돌연변이체들은 제한온도 $42^{\circ}C$에서 고체배지 및 액체배지의 성장은 허용온도에서 배양한 결과보다 저조한 성장을 나타내었다. 감온성 돌연변이체들은 $42^{\circ}C$에서 12시간 배양한 후 허용온도인 $30^{\circ}C$ 액체배지로 이식하여 배양한 결과 치사군중 ts-U1112, ts-D1290, ts-D1342, ts-D1504 돌연변이체들은 성장을 계속하지 않았으며, ts-U351, ts-U828, ts-U892, ts-D1350, ts-D1479, ts-D2601과 ts-D2801 돌연변이체들은 성장이 계속됨을 관찰할 수 있었다.

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A pheromone mutant of Schizosaccharomyces pombe displays nucleolar fragmentation

  • Jun, Jai-Hyun;Kim, Dae-Myung
    • BMB Reports
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    • 제41권3호
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    • pp.248-253
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    • 2008
  • Stresses and nutritional starvation are two main external signals for the induction of sex pheromones in the fission yeast Schizosaccharomyces pombe. In an attempt to identify the components involved in transduction of starvation signals, we screened 135 temperature-sensitive (ts) mutants and isolated 6 mutants that induced the pheromone even in the presence of a nitrogen source. These mutants exhibited two distict induction phenotypes: pheromone induction at restrictive but not at permissive temperatures; and pheromone induction at both permissive and restrictive temperatures. The times required for the maximum pheromone induction at the restrictive temperature differed slightly in each mutant. In addition to the pheromone induction phenotype, the ts243 and ts304 mutants exhibited cell-division-cycle defects. The ts304 mutant cells showed an abnormal cytoplasmic DAPI staining pattern. The nucleolus of this mutant seemed to be fragmented, a phenomenon which is typically observed in aged yeast cells. The result of our genetic analysis indicated that the pheromone induction mutants belonged to 6 separate complementation groups. We designated these mutants pws1 to pws6.

Bacillus thuringiensis serovar kurstaki 돌연변이 균주의 편모항원의 변이성 (Immunological Variations of Flagella Antigens in Bacillus thuringiensis serovar kurstaki Temperature-sensitive Mutants)

  • 이형환;이영주;이희무
    • 한국미생물·생명공학회지
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    • 제17권1호
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    • pp.81-83
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    • 1989
  • The flagella antigenic variation of nine Bacillus thuringiensis serovar kurstaki temperature-sensitive mutants grown at the permissive temperature (3$0^{\circ}C$) was detected by a serological agglutination between H-antigen and antiserum. The flagella antigens were injected to rabbits to prepared their antisera, and then their homologous and heterologous titers of the antisera were measured. The homologous titers were ranged from 1:6,400 to 1:12,800, but the heterologous titers were very low. The H-antigen of the wild type strain was not agglutinated to 4 heterologous antisera, ts-U23 not to 7, ts-U3l not 5, ts-U32 not to 4, ts-U33 not to 7, ts-U7l not to 4, ts-U73 not to 6, ts-U74 not to 6, ts-U91 not to 4 and ts-U603 not to 4 antisera.

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Bacillus thuringiensis 의 Temperature-sensitive Mutants 분리와 특성 연구 (Growth Patterns of Temperature-sensitive Mutants of Bacillus Thuringiensis)

  • Lee, Hyung-Hoan;Lee, Hoon-Ku
    • 한국미생물·생명공학회지
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    • 제11권3호
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    • pp.233-239
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    • 1983
  • Bacillus thuringiensis 3ab K- 3 균주를 28$^{\circ}C$에서 UV광으로 돌연변이를 유도한 다음에 42$^{\circ}C$에서 24개의 Temperature-sensitive(ts)돌연변이 균주를 분리하여 42$^{\circ}C$에서의 성장과 항생제 저항성에 대한 성격을 연구하였다. 24개의 ts 돌연변이 균주중에 13개의 ts돌연변이체는 42$^{\circ}C$의 제한온도에서 전혀 성장을 하지 않았으며, 나머지 11개의 돌연변이체는 성장이 28$^{\circ}C$에서 보다 1/2 에서 1/4까지 감축되었다. 이러한 성격에 따라서 ts돌연변이체는 치사군과 비 치사군으로 나눌수 있었다. 또한 B. thuringiensis SI K-2을 nitrosoguanidine으로 28$^{\circ}C$에서 처리하여 7개의 Auxotrophic mutant을 분리하였다. A-N28은 Arg$^{-}$ . Gly$^{-}$ , Leu$^{-}$ , Met$^{-}$ , Ser$^{-}$ , Val$^{-}$ 을 요구했고, A-N 65는 Ala $^{-}$ , Gly$^{-}$ , Leu$^{-}$ , Met$^{-}$ , Ser$^{-}$ , Val$^{-}$ 을 요구했으며 A-N 92는 Gly$^{-}$ , Leu$^{-}$ , Met$^{-}$ 을, A-N115는 7개의 아미노산을 A-N156은 8개를 요구했다. B. thuringiensis 3ab K-3와 SI K-2 는 Penicillin, Ampicillin과 Cephalothin에 저항성을 나타냈고 ts-U171, A-N92와 A-N115는 이 세항생제에 민감성을 나타냈다. Temperature - sensitive돌연변이균주를 42$^{\circ}C$에서 24시간 배양후에 28$^{\circ}C$로 옮기어 배양한 결과 ts-U601, -U603, -U604와 -U171은 성장을 더 이상하지 않았다.

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Aspergillus nidulans 온도감수성 돌연변이주의 분리 및 분석 (Isolation and Analysis of Temperature Sensitive Mutants in Aspergillus nidulans)

  • 박찬규;강현삼
    • 미생물학회지
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    • 제16권3호
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    • pp.111-121
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    • 1978
  • About 40 temperature-sensitive mutants have been isolated as a preliminary step to study the spore germination, the cell cycle, and the control of macromolecular synthesis in Aspergillus nidulans. To obtain temperature-sensitive mutants rapidly and effectively, the selective enrichment method using antifungal antibiotic nystatin was developed. Based on the data which had applied to the concentration of auxotrophic mutants by the earlier investigators, the optimal concentration and the time of treatment at the nonpermissive temeprature were determined as 50 to 100 units per ml and 4.5 hr., respectively. Out of 41 ts mutants assigned to the strain symbol PK, thirteen that seemed to be arrested at the earlystage of spore germination were subjected to the further cytological and genetic analysis. Elght of these mutants are able to form germ tube and five not. Staining with acid fuchsin for the 5PK strains shows that one irreversible mutant, PK6 strain able to form germ tube, accumulate mitotic spindle, being arrested in mitosis. Another PK15 and PK23 strain have more than one intact nucleolus without germ tube formation at the restrictive temperature. the temperature-senstive mutation in PK12 strain, the onlystrain which is able occurred in certain gene specific for the germination of spore. All of the ts markers are recessive and complement each other in heterokaryon between two different ts markers at the restrictive temperature.

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Identification of virulence-associated genes of Erwinia amylovora by transposon mutagenesis

  • Seung Yeup Lee;Hyun Gi Kong;In Jeong Kang;Hyeonseok Oh;Hee-Jong Woo;Eunjung Roh
    • 농업과학연구
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    • 제50권2호
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    • pp.241-247
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    • 2023
  • Erwinia amylovora , which causes fire blight disease on apple and pear trees, is one of the most important phytopathogens because of its devastating impact. Currently, the only way to effectively control fire blight disease is through the use of antibiotics such as streptomycin, kasugamycin, or oxytetracycline. However, problems with the occurrence of resistant strains due to the overuse of antibiotics are constantly being raised. It is therefore necessary to develop novel disease control methods through an advanced understanding of the pathogenesis mechanism of E. amylovora . To better understand the pathogenesis of E. amylovora , we investigated unknown virulence factors by random mutagenesis and screening. Random mutants were generated by Tn5 transposon insertion, and the pathogenicity of the mutants was assessed by inoculation of the mutants on apple fruitlets. A total of 17 avirulent mutants were found through screening of 960 random mutants. Among them, 14 mutants were already reported as non-pathogenic strains, while three mutants, TS3128_M2899 (ΔSUFU ), TS3128_M2939 (ΔwcaG ), and TS3128_M3747 (ΔrecB ), were not reported. Further study of the association between E. amylovora pathogenicity and these 3 novel genes may provide new insight into the development of control methods for fire blight disease.

Salmonella enterica serovars Enteritidis의 온도감수성 변이주 및 폴리인산키나아제 변이주의 제작과 방어효과 (Development and evaluation of protective capacity of Salmonella Enteritidis polyphosphate kinase-deleted and temperature-sensitive mutant)

  • 김기주;박소연;조영재;곽정연;강정무;김은희;최환원;원호근;노윤희;한태욱
    • 대한수의학회지
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    • 제53권4호
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    • pp.211-216
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    • 2013
  • This study was focusing on evaluating the protection of polyphosphate kinase (ppk) deleted and/or temperature-sensitive (ts) Salmonella Enteritidis (SE) as an attenuated vaccine in chickens. We constructed SEppk, SEts and SEppk::ts mutants and screened those mutants by growth capability in vitro, protection study in mice model and antibody response in chickens. Among the mutants, SEppk::ts-3 was selected because it showed higher growth capability, good protection against highly virulent SE in mice model, and good antibody response in chickens. SEppk::ts-3 also showed good protection against highly virulent SE isolate because it decreased colonization of virulent SE challenge strain in spleen, liver and cecum compared with the non-vaccinated control. The SEppk::ts-3 mutant showed cross-protection against S. Gallinarum (SG) challenge although the its cross-protection rate was a little lower than that of SG9R, a commercial vaccine against SG infection. To use for live attenuated vaccine in chickens, it should further be characterized.

Isolation and Characterization of Pre-$tRNA^{Val}$ Splicing Mutants of Schizosaccharomyces pombe

  • Hwang, Ku-Chan;Kim, Dae-Myung
    • Journal of Microbiology
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    • 제35권4호
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    • pp.334-340
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    • 1997
  • A collection of 132 temperature sensitive (ts) mutants was generated by the chemical mutagenesis of Schizosaccharomyces pombe wild type strain and screened for tRNA splicing defects on Northern blots by hybridization with an oligonucleotide that recognizes the exon of the S. pombe tRNA^Val as a probe. We identidied 6 mutants which accumulate precursor $tRNA^{Val}$. Among them, 2 mutants exhibited remarkable morphological differences compared to wild type cells. One tRNA splicing mutant showed elongated cell shape in permissive as well as non-permissive cultures. The other mutant exhibited shortened cell morphology only in nonpermissive culture. The total RNA pattern in the splicing mutants appeared to be normal. Genetic analysis of four $tRNA^{Val}$ splicing mutants demonstrated that the mutation reside in different genes.

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