• 제목, 요약, 키워드: Synechocystis PCC 6803

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Synechocystis sp. PCC 6803으로부터 광활성 종속영양 생장결핍 돌연변이체의 분리 및 특성 (Isolation and Characterization of a Mutant Defective in Light-activated Heterotrophic Groth from Synechocystis sp. PCC 6803)

  • 박미선;이영숙;김영창
    • 미생물학회지
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    • v.32 no.3
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    • pp.202-207
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    • 1994
  • Bacillus megaterium ATCC 14945의 penicillin G acylase 유전자의 염기배열을 결정하였다. 이 유전자에는 2,406 염기쌍으로 이루어진 하나의 open reading frame이 존재하는데, 개시코돈의 5' 위쪽에서 Shine-Dalgarno 배열과 promoter로 여겨지는 부분을 발견하였으며, 종결코돈의 3' Synechocystis sp. PCC 6803으로부터 광활성종속영양으로 생장할 수 없는 돌연변이체 PRM1을 분리하였다. PRM1을 혼합영양으로 배양하였을 경우에는 생장속도가 Synechocystis 6803과 거의 같았다. 그러나 PRM1을 하루에 5 분만 빛을 조사하면서 광활성종속영양으로 배양하였을 경우에는 전혀 생장하지 못하였다. 이러한 결과는 PRM1이 종속영양으로 생장하는데 필요한 대사능력에 이상이 있는 것이 아니라 생장에 필요한 광신호 전이 체계에 이상이 있음을 시사한다. PRM1의 plasmid 양상, 균체의 absorption spectra, 세포 내부와 외부의 형태 등은 Synechocystis 6803과 유사하였으나 여러 세포들을 함께 얽어매는 부정형 점액성 물질을 형성하지 않는 점이 달랐다.

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Two-component Signal Transduction in Synechocystis sp. PCC 6803 under Phosphate Limitation: Role of Acetyl Phosphate

  • Juntarajumnong, Waraporn;Eaton-Rye, Julian J.;Incharoensakdi, Aran
    • BMB Reports
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    • v.40 no.5
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    • pp.708-714
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    • 2007
  • The two-component signal transduction, which typically consists of a histidine kinase and a response regulator, is used by bacterial cells to sense changes in their environment. Previously, the SphS-SphR histidine kinase and response regulator pair of phosphate sensing signal transduction has been identified in Synechocystis sp. PCC 6803. In addition, some response regulators in bacteria have been shown to be cross regulated by low molecular weight phosphorylated compounds in the absence of the cognate histidine kinase. The ability of an endogenous acetyl phosphate to phosphorylate the response regulator, SphR in the absence of the cognate histidine kinase, SphS was therefore tested in Synechocystis sp. PCC 6803. The mutant lacking functional SphS and acetate kinase showed no detectable alkaline phosphatase activity under phosphate-limiting growth conditions. The results suggested that the endogenous acetyl phosphate accumulated inside the mutants could not activate the SphR via phosphorylation. On the other hand, exogenous acetyl phosphate could allow the mutant lacking functional acetate kinase and phosphotransacetylase to grow under phosphate-limiting conditions suggesting the role of acetyl phosphate as an energy source. Reverse transcription PCR demonstrated that the transcripts of acetate kinase and phospho-transacetylase genes in Synechocystis sp. PCC 6803 is up-regulated in response to phosphate limitation suggesting the importance of these two enzymes for energy metabolism in Synechocystis cells

Optimum Conditions for Transformation of Synechocystis sp. PCC 6803

  • Zang, Xiaonan;Liu, Bin;Liu, Shunmei;Arunakumara, K.K.I.U.;Zhang, Xuecheng
    • Journal of Microbiology
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    • v.45 no.3
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    • pp.241-245
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    • 2007
  • This study was conducted to determine the optimal conditions for introduction of exogenous DNA into Synechocystis sp. PCC 6803. Of the three transformation techniques studied, electroporation, ultrasonic transformation and natural transformation, natural transformation showed the highest efficiency. Additionally, this study demonstrated that the higher plasmid concentration and longer homologous recombining fragments resulted in a greater number of transformants. For successful transformation, the lowest concentration of plasmid was $0.02\;{\mu}g/ml$, and the shortest homologous recombining fragment was 0.2 kb. Use of Synechocystis sp. PCC 6803 in the logarithmic growth phase resulted in two-fold higher transformation rate than that of the same organism when cells in the latent phase or the plateau phase were used for transformation. Pretreatment of the host strain, Synechocystis sp. PCC 6803, with EDTA (2 mM) for two days prior to transformation increased the transformation efficiency by 23%. Additionally, incubation of the cells and DNA for 5 h under light conditions increased the transformation efficiency by two orders of magnitude. Moreover, recovery treatment of the cells before they were plated onto antibiotic medium also increased the transformation efficiency.

중금속이 Cyanobacterium synechocystis sp.PCC 6803의 성장과 단백질 합성에 미치는 영향 (Effects of Heavy Metals on Growth and Protein Synthesis in Cyanobacterium synechocystis sp. PCC 6803)

  • 강경미;장남기
    • 아시안잔디학회지
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    • v.10 no.4
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    • pp.315-329
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    • 1996
  • The changes of growth and protein synthesis pattern by aluminum (Al), cadmium (Cd), zinc (Zn) treatments were studied in Cyanobacterium synechocystis sp. PCC 6803. When exposed to Al from 5ppm to 3oppm, synechocystis grows normally. But more than that retard the growth of algae notably. The 0.05ppm Cd additions had no effect on the growth of algae. 0.1, 0.2, and 0.5ppm Cd inhibited growth. Under 1 and 2ppm Cd stress, growth was greatly diminished. Zn had dual effects. The growth of algae in media containing 5ppm was stimulated. As concentration increases more than l5ppm, growth inbition increases. Under 25ppm Zn stress, growth was greatly diminished. According to logistic theory, r and K values of each heavy metal-treated groups were estimated. Correlation analysis of r and K values with metal concentration shows that there is negative correlation between K and concentration in Cd and Zn treatments. Critical concentration which shows lethal or sublethal effect was estimated by t-test of each r and K value. The cells cultured in 10, 20, 30, 40 and 5oppm of Al, 1 and 2ppm of Cd, and 10, 15, 20, 25 and 30ppm of Zn for 4 days was used for protein analysis. Analysis of protein synthesis with SDS-PACE showed alterations of protein synthesis pattern. The synthesis of protein about 220kD increased markedly. In this study, it showed that resistance mechanism against Al, Cd, and Zn is K selection and that metal stress induced the change of protein synthesis in Cyanobacterium synechocystis sp. PCC 6803.Key words:Cyanobacterium synechocystis sp. FCC 6803, Heavy metals, Aluminum, Cadmiutm Zinc, Crowth, Frotein synthesis.

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Synechocystis sp. PCC6803을 이용한 Photosystem I- mutants의 색소 및 틸라코이드막 단백질 분석 (Analysis of Pigments and Thylakoid Membrane Proteins in Photosystem I - Mutants from Synechocystis sp. PCC6803)

  • 전은경;장남기
    • 아시안잔디학회지
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    • v.11 no.1
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    • pp.45-58
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    • 1997
  • Pigments and thylakoid membrane proteins were investigated in wild type and PS I- mutants from Synechocystis sp. PCC6803 Comparing morphological features, B2 was less fluorescent than the other strains. The contents of chlorophyll a were propotional to the FNR activity in thylakoid membrane. The FNR activity of mutants was lower than that of wild type. In the result of pigments analysis, mutants had smaller cholophyll a than that of wild type. The major carotenoid was found to he $\beta$-caroene, but aeaxanthin was barely detected in thylakoid membrane of mutants. The polypeptide, 14.8kD was detected by electrophoresis in mutants. It was considered to be the modification of 15.4kD in wild type. Membrane polypeptides of 17.6 and 19.7kD were not detected in mutants. In the result of western blotting, subunit I was detected in all strains, but subunit II was barely detected in mutants. Subunit II was not detected in B2 at all. In view of the results so far achieved, the changes of contents of chlorophyll and zeaxanthin were affected by the defficiency or modification of functional domain in subunit I. Also the modification in subunit I affected the subunit II- binding site in PS I. As the result, efficiency of photosynthesis was decreased. Key words: Synechoystis sp. PCC6803, PS I - mutant, Photosynthetic efficiency, Pigment,Thylakoid membrane proteins, Subunit I, II.

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Bacillus thuringiensis subsp. morrisoni PG-14 cryIVD 유전자로 형질전환된 Synechocytis PCC6803의 특성과 학질모기에 대한 살충효과 (Characterization of Synechocystic PCC6803 transformed with cryIVD gene of Bacillus thuringiensis subsp. morrisoni PG-14 and its mosquitochidal effect on Anopheles sinensis)

  • 이대원;박현우;김호산;진병래;유효석;김근영;강석권
    • 한국응용곤충학회지
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    • v.35 no.1
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    • pp.66-73
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    • 1996
  • Bacillus thuringiensis subsp. morrisoni PG-14의 cryIVD 유전자를 포함하고 있는 발현벡터 pCYASK5-1을 제작하여 Synechocystis PCCC6803에 형질전환시킨 후, 학질모기(Anopheles sinensis)유충에 대한 독성을 검정하였다. Kanamycin이 포함된 BG-11배지에서 선발된 형질전환체의 cryIVD 유전자 발현은 SDS-PAGE와 West-ern blot분석으로 확인하였다. 형질전환체는 A. sinensis 유충에 대해 높은 독성을 나타내었으며, 성장은 야생주인 Synechocystis Pccc6803과 유사하였다. 또 수심에 따른 형질전환체의 분포도 조사에서 전체적으로 살충농도의 세포수로 분포함을 확인하였다. 이러한 결과들은 B. thuringiensis subsp. morrisoni PG-14의 cryIVD 유전자로 형질전환된 Synechocystis PCCC6803이 모기유충 방제에 효율적으로 이용될 수 있음을 나타내었다.

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Cyanobacterium Synechocystis sp.PCC6803 psaB 돌연변이주의 광합성 전자전달에 미치는 호흡의 영향 (Influence of Respiration on Photosynthetic Electron Transport in psaB Mutants from Cyanobacterium Synechocystis sp. PCC6803)

  • 윤병철;장남기
    • 아시안잔디학회지
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    • v.11 no.1
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    • pp.59-72
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    • 1997
  • The influence of respiration on photosythetic electron transport were investigated in the Wid type and psaB mutants from Syneehocystis sp. PCC6803. The amount of glucose uptake in the wild type was proportional to the glucose concentration added in wild type and less than that of psaB mutants in the dark. It was suggested that psaB mutants more depend on the glucose than the wild type. It was investigated how the activities of isocitrate dehydrogenase(IDH) and glucose-6-phos-phate dehydrogenase(G6PDH) were changed. The activities of IDH were very low. While, the ac-tivities of G6PDH were much higher than that of IDH. These results agree to the reports that ex-ogenous glucose was dismilated aerobically via Oxidative Pentose Phosphate Pathway in heterotrophic cyanobacteria. PsaB mutants showed high G6PDH activity in the presence of glucose as well as in the dark and high respiratory activities especially in the dark. It was also investigated how photosynthetic electron transport activities were changed. PsaB mutants showed higher photosynthetic electron tranasport activities than wild type in the presence of glucose as well as in the dark. In the results, it was proposed that photosynthetic electron transport between PS I and PS U was complemented by respiratory electron transport through the NADPH generated by Dxidative Pentose Phophate Pathway in psaB mutant from Synechocystis sp. PCC6803. Key words: Photosynthetic electron transport, Respiration, Synechoystis sp. PCC6803, psaB mutant, Glucose uptake, IDH, G6PDH, Respiratory electron transport activity.

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Cryo-Methods와 Cryo-SEM을 이용한 Cyanobacteria, Synechocystis sp. PCC 6803 미세구조 관찰 (Observations of the Cyanobacteria Synechocystis sp. PCC 6803 using Cryo-Methods and Cryo-SEM)

  • 이은주;문윤정;오현우;김수진;정영호;권희석;김윤중
    • Applied Microscopy
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    • v.39 no.1
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    • pp.65-72
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    • 2009
  • Cryo-SEM은 수분을 함유하거나 액상 시료를 동결된 상태로 관찰하는 방법으로 rapid cooling, fracturing, sectioning, etching, coating과 같은 다소 복잡한 여러 과정의 전처리(Cryo-methods) 로 구성된다. 각 과정에서는 분석 목적이나 시료 특성(예: 시료크기, 물 함량 등)을 고려하여 최적의 조건을 설정하여야 한다. 본 연구에서는 cryo-SEM을 이용하여 남세균 (cyanobacteria, Synechocystis sp. PCC 6803)의 표면 및 내부 구조 관찰을 위한 etching time과 시료 처리에 관하여 살펴보았고 cryo-methods로 처리한 후 cryo-SEM으로 관찰한 이미지를 화학 고정한 일반 SEM(CSEM) 결과와 비교하였다. 관찰 결과, 화학 고정한 CSEM에서는 Synechocystis sp. PCC 6803 표면에 존재하는 pili가 잘 관찰되지 않았으며 파단된 내부 구조의 이미지를 얻을 수 없었으나 cryo-methods/cryo-SEM에서는 세포 표면의 pili 및 membrane의 형태를 관찰할 수 있었다. 또한 수화 상태에 따라 구조변형이 일어나는 biofilm의 구조도 관찰할 수 있었다. 이러한 결과로부터 cryo-methods/cryo-SEM은 수분을 함유하는 의생물 시료의 형태 구조를 분석하는 유용한 방법으로 사료된다.