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Browsing Graduate School by Title "<i>Karenia</i> species in the Gulf of Mexico: Cell Cycle Characterization and Development of a Novel Tool for Species Specific Protein Analysis"
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- ItemKarenia species in the Gulf of Mexico: Cell Cycle Characterization and Development of a Novel Tool for Species Specific Protein AnalysisMeek, MeganHarmful algal blooms of Karenia species have been documented in the Gulf of Mexico for over a century. At least four species are known to co-occur in blooms: Karenia brevis, Karenia mikimotoi, Karenia papilionacea, and Karenia selliformis. The first objective of this study was to characterize the cell cycle progression of each Karenia species, and examine their expression of the cell cycle-specific protein proliferating cell nuclear antigen (PCNA) over the diel cycle. This work found that all four species exhibited diel phasing of their cell cycles, and observed a strong significant relationship between PCNA and S-phase of the cell cycle in K. brevis, K. mikimotoi and K. selliformis (p ≤ 0.01), as well as a less significant relationship in K. papilionacea (p < 0.10) These results indicate the potential for the use of PCNA in a species-specific growth assay. The second objective of this study further examined the relationship between PCNA abundance and growth rates in K. brevis using flow cytometry to quantify PCNA abundance, but found no significant relationship, possibly due to the differing epitope cross-reactivity of a newly synthesized PCNA antibody. A dual labeling method was designed combining a K. brevis specific fluorescent in-situ hybridization (FISH) probe and the PCNA antibody to analyze intracellular protein in a species-specific method. This is the first report of such a dual labeling technique in a phytoplankton species, and the method has many potential applications for phytoplankton research. Its utility in Karenia species, for species-specific growth rate determination remains viable, but would require a more effective PCNA antibody.
- ItemKarenia species in the Gulf of Mexico: Cell Cycle Characterization and Development of a Novel Tool for Species Specific Protein AnalysisMeek, MeganHarmful algal blooms of Karenia species have been documented in the Gulf of Mexico for over a century. At least four species are known to co-occur in blooms: Karenia brevis, Karenia mikimotoi, Karenia papilionacea, and Karenia selliformis. The first objective of this study was to characterize the cell cycle progression of each Karenia species, and examine their expression of the cell cycle-specific protein proliferating cell nuclear antigen (PCNA) over the diel cycle. This work found that all four species exhibited diel phasing of their cell cycles, and observed a strong significant relationship between PCNA and S-phase of the cell cycle in K. brevis, K. mikimotoi and K. selliformis (p ≤ 0.01), as well as a less significant relationship in K. papilionacea (p < 0.10) These results indicate the potential for the use of PCNA in a species-specific growth assay. The second objective of this study further examined the relationship between PCNA abundance and growth rates in K. brevis using flow cytometry to quantify PCNA abundance, but found no significant relationship, possibly due to the differing epitope cross-reactivity of a newly synthesized PCNA antibody. A dual labeling method was designed combining a K. brevis specific fluorescent in-situ hybridization (FISH) probe and the PCNA antibody to analyze intracellular protein in a species-specific method. This is the first report of such a dual labeling technique in a phytoplankton species, and the method has many potential applications for phytoplankton research. Its utility in Karenia species, for species-specific growth rate determination remains viable, but would require a more effective PCNA antibody.