Team:Kyoto/Result

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1) Salt concentration has various effects on protein-protein interaction

In starting the experiment, we conducted an experiment demonstrating how the salt concentration of the solution actually affects the protein-protein interaction by using the model protein. In this example, we chose GFP BBa_E0040, which is commonly used in many experiments as a model protein.
TDH3 promoter and CYC1 terminator were added to both ends of ORF and cloned into pRS316 which is a shuttle vector of S. cerevisiae and E. coli. The resulting plasmid was transformed into wild-type yeast strain BY4741 to overexpress GFP in yeast. As a comparative experiment, yeast expressing RFP (BBa - E 0010) with the same set of promoter and terminator was used.
First, photographs of pellets recovered from the culture medium of yeast cells used in this experiment are shown.

Figure1.A picture of yeast expresses RFP or GFP

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Reference

• [1] X. rong Wang, X. Cheng, Y. dong Li, J. ai Zhang, Z. fen Zhang, and H. rong Wu, “Cloning arginine kinase gene and its RNAi in Bursaphelenchus xylophilus causing pine wilt disease,” Eur. J. Plant Pathol., vol. 134, no. 3, pp. 521–532, 2012.
• [2] A. Sigova, N. Rhind, and P. D. Zamore, “A single Argonaute protein mediates both transcriptional and posttranscriptional silencing in Schizosaccharomyces pombe,” genes Dev., 2004.
• [3] R. Esteban and R. B. Wickner, “A new non-mendelian genetic element of yeast that increases cytopathology produced by M1 double-stranded RNA in ski strains.,” Genetics, 1987.
• [4] M. T. B. Sloan, Katherine E, Pierre-Emmanuel Gleizes, “Nucleocytoplasmic Transport of RNAs and RNA–Protein Complexes,” J. Mol. Biol., vol. 428, no. 10, pp. 2040–2059, 2016.
• [5] V. W. Pollard and M. H. Malim, “the Hiv-1 Rev Protein,” Annu. Rev. Microbiol., vol. 52, no. 1, pp. 491–532, 1998.