Graduate Thesis Or Dissertation
 

Exogenous gene expression from heterologous promoters in fish cell cultures

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  • Cell culture systems have provided many insights into eukaryotic gene expression and other biochemical mechanisms. Since the cell represents the smallest living unit of any organism it provides a desirable in vitro system, allowing biochemical studies without the complex physiology of an entire animal. However, processes involving intracellular mechanisms, such as development, aging or carcinogenis, eventually require the analysis of the intact organism. Transgenic animals are a very promising tool to approach questions of this magnitude. Fish in general and the zebrafish (Brachydanio rerio) in particular are an excellent model system for transgenic research, mainly due to their extramaternal fertilization and development and their short generation cycle throughout the year. The recent derivation of zebrafish cell lines has opened up possibilities for in vitro analysis of this popular model species, and expression of heterologous genes under the influence of promoter and other regulatory nucleic acid aequences. In contrast to mammalian expression systems, little nucleic acid sequences controlling gene expression in fish are known. Therefore we examined mammalian expression systems in fish cells in order to determine their efficiency quantitatively. Emphasis was given to zebrafish cultures with the goal of eventually injecting in vitro manipulated embryo cells into host embryos and thereby creating transgenic chimera.
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