• Title, Summary, Keyword: Synechocystis PCC 6803

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

  • Park, Mi-Seon;Lee, Young-Sook;Kim, Young-Chang
    • Korean Journal of Microbiology
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    • v.32 no.3
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    • pp.202-207
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    • 1994
  • A mutant strain PRM1 defective in light-activated heterotrophic growth was isolated from Synechocystis sp. PCC 6803. PRM1 could be grown at growth rate equivalent to Synechocystis 6803 under mixotrophic growth conditions. However, PRM1 could not be grown under light-activated heterotrophic conditions, in which a daily pulse of light for 5 min was given. These results suggest that PRM1 is not defective in heterotrophic metabolism, but in the transduction pathway of light signal essential to the growth. Plasmid patterns, absorption spectra of whole cells, and the exterior and interior structures of PRM1 were similar to those of Synechocystis 6803, except that PRM1 could not produce amorphous slime holding cells together.

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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.

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

  • 강경미;장남기
    • Asian Journal of Turfgrass Science
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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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Analysis of Pigments and Thylakoid Membrane Proteins in Photosystem I - Mutants from Synechocystis sp. PCC6803 (Synechocystis sp. PCC6803을 이용한 Photosystem I- mutants의 색소 및 틸라코이드막 단백질 분석)

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

  • 이대원;박현우;김호산;진병래;유효석;김근영;강석권
    • Korean journal of applied entomology
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    • v.35 no.1
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    • pp.66-73
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    • 1996
  • For the effective control of mosquito larvae, Anopheles sinensis, the expression vector pCYASK5-1 containing cryIVD gene of Bacillus thuringiensis subsp. morrisoni PG-14 was constructed and transformed into the cyanobacterium Synechocystis PCC6803. The transformants were selected on BG-11 medium containing kanamycin. The expression of cryIVD gene in transformant was confirmed by SDS-PAGE and Western blot analysis. The mortality of A. sinensis larvae was scored for 3 days. Furthermore, growth and distribution rate of transformant were examined. The results showed that Synechocystis PCC6803 transformed with cryIVD gene of B. thuringiensis subsp. morrisoni PG-14 was highly toxic to A. sinensis larvae, demonstrating that it will be a potential agent for mosquito control.

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

  • 윤병철;장남기
    • Asian Journal of Turfgrass Science
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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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Observations of the Cyanobacteria Synechocystis sp. PCC 6803 using Cryo-Methods and Cryo-SEM (Cryo-Methods와 Cryo-SEM을 이용한 Cyanobacteria, Synechocystis sp. PCC 6803 미세구조 관찰)

  • Lee, Eun-Ju;Moon, Yoon-Jung;Oh, Hyun-Woo;Kim, Su-Jin;Chung, Young-Ho;Kweon, Hee-Seok;Kim, Youn-Joong
    • Applied Microscopy
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    • v.39 no.1
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    • pp.65-72
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    • 2009
  • Cryo-SEM which enables specimens to be observed in frozen form has been used to study liquid specimens in their native states. Cryo-methods, sample preparation for cryo-SEM, are quite complex and involve several discrete but vitally interconnected steps which are rapid cooling, fracturing, sectioning, etching and coating. It is important to select practical techniques and to optimize conditions of each steps considering analytic purpose and specimen characters, viz., sample dimension, water contents. In this study, etching methods and sample preparation before freezing had been studied for observation of cyanobacteria, Synechocystis sp. PCC 6803 using cryo-SEM and their cryo-SEM images were compared to Conventional SEM (CSEM) images treated by chemical fixation. We could observe the improved morphological images of the pili of the surface and membranes of Synechocystis sp. PCC 6803 and the three-dimensional architectures of their biofilm, which were difficult to observe using chemical fixation and CSEM. These results suggest that cryo-methods/cryo-SEM are useful techniques for morphological study of biological specimen.