Article
Advanced method for high-throughput expression of mutated eukaryotic membrane proteins in Saccharomyces cerevisiae.
Biochemical and biophysical research communications - 11 Jul 2008
Ito Keisuke, Sugawara Taishi, Shiroishi Mitsunori, Tokuda Natsuko, Kurokawa Azusa, Misaka Takumi, Makyio Hisayoshi, Yurugi-Kobayashi Takami, Shimamura Tatsuro, Nomura Norimichi, Murata Takeshi, Abe Keiko, Iwata So, Kobayashi Takuya
Abstract excerpt
Crystallization of eukaryotic membrane proteins is a challenging, iterative process. The protein of interest is often modified in an attempt to improve crystallization and diffraction results. To accelerate this process, we took advantage of a GFP-fusion yeast expression system that uses PCR to direct homologous recombination and gene cloning. We explored the possibility of employing more than one PCR fragment to...
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