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Identifier 000388636
Title Απομόνωση και πρωτεομική ανάλυση των ετεροκυστών από το κυανοβακτήριο Anabaena sp. PCC 7120
Alternative Title Isolation and proteomic analysis of heterocysts from cyanobacteria Anabaena sp.PCC7120.
Author Μαναρωλάκη, Ευτυχία Σ
Thesis advisor Τσιώτης, Γεώργιος
Reviewer Γανωτάκης, Δημήτριος
Κοτζαμπάσης, Κυριάκος
Abstract Anabaena sp.PCC7120 is a Gram negative cyanobacterium, which is able to perform oxygenic photosynthesis and nitrogen fixation. The main characteristic of this bacterium is the complexity of the cell due to the growth of two types of cells (vegetative cells and heterocysts). Specifically, the resistant cell types heterocysts, in nitrogen starvation conditions, produce molecular hydrogen. To investigate the differentiation or not the cells into heterocysts during deprivation was performed by the grow of bacterium for five days in medium BG11 and then two more days in medium BG11o.The present study focused on the implementation of an isolation protocol of heterocysts and to execute two methods for proteomic study. With the first method was achieved selective solubilization of the inner membrane of isolated heterocysts with anionic surfactant, for four different strains of the bacterium (WTPCC7120, hupL-, hoxH-, hupL- / hoxH-). For the analysis and characterization of the fragments obtained, the soluble and insoluble, applied dimensional Tricine SDS PAGE electrophoresis and identified the protein spots which were distinguished by mass spectrometry. Differences in the expression of proteins in different strains was evident. Were identified basic proteins of metabolic pathways for the development and survival of heterocysts such as Alr2269, Alr4550 and Alr2887 with a higher percentage in the strain which is not genetically modified. The number of proteins decreases for other strains and so we can observe the expression of unique proteins in them. By the second method analyzed proteomic, whole heterocysts without prior rupture of outer membranes or the thylakoid membranes and the cytoplasmic membrane. The two-dimensional electrophoresis of isolated bacterium heterocysts was performed by 2D IEF / SDS-PAGE. The procedure showed the successful separation of membrane proteins, since a satisfactory percentage were identified. Additional with the second method identified proteins involved in the differentiation of vegetative cells to heterocysts as in the first method. This allowed us to estimate that the isolation was achieved to some extent and thus to understand better their metabolism and the proteomic composition and operation of a single outer membrane of both cell types. Additionally, we have investigated the hydrogen production from the cell strains and we observe a significantly higher amount of strain hupL-.
Language Greek
Subject Ετεροκύστες
Issue date 2014-11-21
Collection   School/Department--School of Sciences and Engineering--Department of Chemistry--Post-graduate theses
  Type of Work--Post-graduate theses
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