| Summary: | 碩士 === 中原大學 === 化學研究所 === 96 === The bi-cistronic vector and restriction enzyme system had been used to study IRES for many years. Both the techniques have been established allow construction of plasmids to express in mammalian cells and insect cells. Nevertheless, it remains some problems in this system, such as the difficulties of construction of bi-cistronic vectors, complicated works for cloning, and finding of suitable restriction enzyme sites. Those hurdles increase the inconvenient works for researchers and cost more time and money. Yeast is a popular biological system in industry and academic researches. The advantages of yeast include the fast growth rate, well-known genomic sequences and many useful selection markers. Thus, yeast vector expression system should be suitable for the IRES research. Gateway cloning is a newly invented “integrase site-specific recombination system” for plasmid expression, which has been used in many kinds of species. The convenience of Gateway system can assistant bi-cistronic system construction to facilitate IRES element tests. To adopt the shuttle vector between yeast and bacterium, we combined Gateway and bi-cistronic system together to do IRES expression in eukaryotic cells. We call it Gateway bi-custronic shuttle vector system.
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