• 제목/요약/키워드: Mutants

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Roles of Glutathione Reductase and $\gamma$-Glutamylcysteine Synthetase in Candida albicans

  • Baek, Yong-Un;Yim, Hyung-Soon;Kang, Sa-Ouk
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2003년도 정기총회 및 학술발표회
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    • pp.61-61
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    • 2003
  • We have cloned the CGR1 gene encoding glutathione reductase (GR) which catalyzes the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH) from Candida albicans. The cgr1/cgr1 mutants were not viable when CaMAL2 promoter repressed the CGR1 expression. The growth of the mutants could be partially overcome by thiol compounds such as GSH, dithiothreitol, cysteine, N-acetylcysteine and GSSG. Interestingly, C. albicans with CGR1 overexpressed showed defective hyphal growth on solid medium and attenuated virulence. We have also cloned the GCS1 gene encoding ${\gamma}$-glutamylcysteine synthetase which catalyzes the first step of glutathione biosynthesis. The gcs1/gcs1 mutants were nonviable in minimal defined medium. The growth of the mutants could be resumed by supplementing with GSH, GSSG and ${\gamma}$-glutamylcysteine in the medium. The mutants had increased intracellular D-erythroascorbic acid level up to 2.25-fold when transferred to GSH-free medium. When the mutants were depleted of GSH, they showed typical markers of apoptosis. In conclusion, these results suggest that glutathione is an essential metabolite, and involved in hyphal growth, virulence and apoptosis in C. albicans.

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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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Aspergillus nidulans에서 NSD 돌연변이주의 분리 및 분석 (Isolation and Characterization of NSD mutants in Aspergillus nidulans)

  • 한동민;한유정;김지현;장광엽;정윤신;정재훈;채건상
    • 한국균학회지
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    • 제22권1호
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    • pp.1-7
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    • 1994
  • Several mutants which never underwent to sexual development(NSD) of Aspergillus nidulans were analyzed genetically and physiologically. They were divided into two groups according to their characterisitics of asexual development after release from aeration block. The mutants in first group proceeded asexual development immediately after removal of aeration block, while those in second group did 10 hours or more later. The NSD mutants were separated into 4 complementation groups, nsdA, nsdB, nsdC and nsdD. The nsdA and nsdD genes were linked to AcrA1 on linkage groupⅡ and pabaA1 on linkage group I, respectively. The mutant alleles were all recessive to wild type allele in heterokayon state. The mutants did not developed cleistothecia on any of carbon sources, except NSD208 which developed cleistothecia on lactose.

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Relationship between Nodulating Characters and Yield Components in Supernodulating Soybean Mutants

  • Park Sei Joon;Youn Jong Tag;Kim Wook Han;Lee Jae Eun;Kwon Young Up;Shin Jin Chul;Seong Rak Chun
    • 한국작물학회지
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    • 제50권1호
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    • pp.5-10
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    • 2005
  • This experiment was conducted to clarify the functions of supernodulating characters on seed yield determination through the comparison of agricultural traits of supernodulating soybean mutants, Sakukei4, SS2-2, and their parent cultivars, Enrei and Shinpaldalkong2. The plant dry weights of supernodulating mutants, Sakukei4 and SS2-2, were $52\%$ and $61\%$ of their wild type parents at full seed stage (R6). However, the relative growth rate (RGR) from the pod set stage (R3) to R6 of Sakukei4 was 0.022 g/g/day and that of SS2-2 was 0.016 g/g/day, which were higher than those of their parents. Nodule number and dry weight were increased in two supernodulating mutants by the R6 stage. The nitrogen concentrations of leaf, petiole and stem of Sakukei4 were higher than those of Enrei. SS2-2 showed higher nitrogen concentration in petiole than Shinpaldalkong2 had. The positive correlations were appeared between nodule dry weight, plant dry weight and pod number, in two supernodulating mutants during the period from R3 to R6 stage. Although all of the yield components and seed yield were lower in two supernodulating mutants than their parents at the stage of full maturity (R8), the harvest index was higher in supernodulating mutants. The increasing rates of pod number to stem dry weight in two supernodulating mutants showed the higher than those of two their parents at R8 stage. In conclusion, the relative growth rates during the early to the middle reproductive growth period were higher in supernodulating mutants than the wild types. This could be resulted in an increase in pod number. The increase of relative growth rate was the result of the successive supplement of nitrogen source from biological nitrogen fixation (BNF) of nodules during the middle reproductive growth period in supernodulating mutants.

Comparison of Resistance Level to Cotton leaf curl virus(CLCuV) Among Newly Developed Cotton Mutants and Commercial Cultivars

  • Akhtar, Khalid P.;Khan, Azeem I.;Hussain, M.;Khan, M.S.I.
    • The Plant Pathology Journal
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    • 제18권4호
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    • pp.179-186
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    • 2002
  • Four newly developed cotton mutants (M-111, M-7662, M-358 and M-218) were compared for their resistance against Cotton leaf curl virus(CLCuV) together with commercial resistant (CIM-443, CIM-482, CIM-473, FH-900 and FH-901) and susceptible (5-12) varieties by artificial inoculation through grafting and under natural field conditions. Infectivity and success of grafting were 100% in all cases. None of the grafted plants were found immune or asymptomatic. All the grafted mutants and most of their single plant progeny rows (SPPRs) showed highly resistant responses as the symptoms displayed by these mutants were milder than the commercial cultivars. Grafted mutants also had delayed disease reactions as they took more time (25-30 days) to produce disease symptoms, as compared with resistant commercial varieties that produced disease 18-22 days after inoculation. Growth of the grafted SPPRs of tested mutants was normal, which is an indication that there will be no production losses. Observations under natural infestation of whitefly showed that two SPPRs of M-ll/CE and M-7662-1/2 and one resistant variety CIM-443 exhibited slight incidence of disease, while one SPPR of M-l1/59 and S-12 were moderately susceptible and highly susceptible with 21% and 97.l% disease incidence, respectively. This study also showed that plants displaying more disease symptoms through grafting were easily infected under natural conditions. These results suggest that preference should be given to those plants that exhibited highly resistant responses after artificial inoculation.

Waterlogging Effects on Nitrogen Accumulation and $N_2$ Fixation of Supernodulating Soybean Mutants

  • Youn, Jong-Tag;Van, Kyu-Jung;Lee, Jae-Eun;Kim, Wook-Han;Yun, Hong-Tae;Kwon, Young-Up;Ryu, Yong-Hwan;Lee, Suk-Ha
    • Journal of Crop Science and Biotechnology
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    • 제11권2호
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    • pp.111-118
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    • 2008
  • Soybean is sensitive to waterlogging stress, leading to reduce their growth and yield significantly. The objective of this study was to characterize the relative sensitivities of biomass accumulation and specific nodule activity under waterlogging stress between supernoduating mutants, 'SS2-2' and 'Sakukei 4' and their wild-type soybeans, 'Sinpaldalkong 2' and 'Enrei', respectively. Flooding treatment was performed to soybean plants grown in a pot by waterlogging for 15 days from the beginning bloom(R1) stage under natural light. The nodule number and weight were considerably decreased by waterlogging stress. The bleeding sap rate of waterlogging soybean plants was decreased by 78-80% in supernodulating mutants and 65-74% in their wild types compared to control plants. The relative ureide-N content was also decreased by waterlogging and the reduction was high in supernodulating mutants. This may cause the massive reduction of shoot and root dry weight and leaf area in waterlogged soybean plants. There was a varietal difference in response to the waterlogging stress. During the waterlogging, supernodulating mutants maintained higher spad value than their wild types. Particularly, the difference between soybean varieties was clear in low rank leaves from the top. Also, supernodulating mutants showed a weak waterlogging tolerance than their wild types. Under waterlogging conditions, massive nodules were considerably destroyed and specific nodule activity after waterlogging may not be recovered when compared to their wild-type soybeans. Supernodulating mutants showed lower seed yield than their wild types in waterlogging conditions.

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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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Heme 합성특성이 다른 Rhizobium meliloti 102F51 Mutant의 선별 (Rhizobium meliloti 102F51 Mutants Defective in Heme Synthesis)

  • 최영주;정원화;김경수;신평균;조무제
    • 미생물학회지
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    • 제24권2호
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    • pp.98-105
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    • 1986
  • Rhizobium melitoli 102F51에 N-methyl-N'-nitro-N-nitrosoguanidine 및 UV를 처리하여 heme 합성 특성이 다른 세 그룹의 mutant 즉 백색, 적백색 및 적색근류형성 mutant를 선별하여 이들 중 mutant들의 acetylene 환원력, 및 ${\delta}-aminolevulinic$ acid dehydratase (ALAD)를 free living 및 bacteriod 상태하에서 서로 비교하였다. 백색 근류를 형성하는 mutant는 적색 근류형성 mutant에 비하여 acetylene 환원력이 훨씬 낮았으며 ALAS 및 ALAD 활성은 free-living 상태에서는 각 mutant group 사이에서 큰 차이를 보이지 않았으나 bacteriod 상태에서는 백색근류생성 mutant에서 적색 근류형성 mutant group에 비하여 ALAS 및 ALAD 공히 현저히 낮았다. 근류에서 ALAS 활성은 heme 합성양에 비례하여 급진적으로 증가하였으나 ALAD 활성은 plant fraction에서는 감소하는 반면 bacteroid fraction에서는 완만하게 증가하였다.

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누에의 후배자발생 초기에 있어서 돌연변이 불면잠계통간의 형질발현 특성 (Pheontypic Characterization of Non-molting Mutants in Postembryonic Development of the Silkworm, Bomyx mori.)

  • 노시갑;선희숙;반야풍
    • 한국잠사곤충학회지
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    • 제40권1호
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    • pp.43-51
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    • 1998
  • 곤충 후배자발생과정(약령기)에 있어서 유전자의 형질발현기구를 구명하기 위한 연구의 일환으로, 누에의 유전적 돌연변이 불면잠계통을 재료로 하여 실험을 실시하였다. nm-d, nm-f, nm-k, nm-i, nm등의 5개 불면잠계통에 대한 불면잠의 발현양상, 생존일수, 치사율, 체액단백질의 농도 및 조성 등을 조사한결과, 불면잠계통들은 대체로 유사한 형질발현을 하였으나 각 계통들의 유전자특유의 형질특성은 인정되었다. 이들 특성을 기초로하여 불면잠계통을 두가지 유형으로 구분할 수 있었다. 즉 치사시기가 빠르고 체액단백질성분의 대부분이 소실되는 약령기퇴화형불면잠과 생존기간이 비교적 길며 체액단백질성분의 감소가 비교적 적은 약령기유지형 불면잠으로 그 특성이 구분되었다.

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Characterization of Insulin-like Growth Factor-free Interaction between Insulin-like Growth Factor Binding Protein 3 and Acid Labile Subunit Expressed from Xenopus Oocytes

  • Choi, Kyung-Yi;Kyung, Yoon-Joo;Lee, Chul-Young;Lee, Dong-Hee
    • BMB Reports
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    • 제37권2호
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    • pp.153-158
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    • 2004
  • The acid-labile subunit (ALS) is known to interact with the IGF binding protein (IGFBP) in the presence of insulin-like growth factors (IGFs). Studies, however, indicate that ALS forms a doublet with IGFBP3, independent of IGFs. To characterize the structural domain required for the IGF-free ALS-IGFBP3 interaction, seven recombinant human IGFBP3 mutants were generated: three deletion mutants and four site-specific mutants that had altering N-terminal regions of IGFBP3. ALS and IGFBP3 mRNAs were co-injected into Xenopus oocytes, and their products were cross-linked and immunoprecipitated using antisera against ALS or IGFBP3. Among the deletion mutants, the mutant of D40 (deleted in 11-40th amino acids) exerted no effect in the interaction with ALS, while D60 (${\Delta}11$-60) demonstrated a moderate reduction. D88 (${\Delta}11$-88), however, showed a significant decrease. In the case of site-specific mutants, the mutation that alterated the IGF binding site (codons 56 or 80) exerted a significant reduction in the interaction, whereas codons 72 or 87 showed no significant change in the interaction with ALS. The stability of the ALS-IGFBP3 interaction was analyzed according to a time-dependent mode. Consistent with the binding study, mutants on the IGF binding sites (56 or 80) consistently show a weakness in the ALS-IGFBP3 interaction when compared to the mutants that covered the non-IGF binding sites (72 or 87). This study suggests that the N-terminal of IGFBP3, especially the IGF binding site, plays an important role in interacting with ALS as well as in stabilizing the dual complex, independent of IGFs.