- 第3日 5月17日（木） 10:45～11:00 A会場（オービットホール）
Proteomic methods using mass spectrometry have been developed by combination of a variety of techniques. Here we introduce our originally developed strategies in combination with different kinds of techniques along with obtained results. Absolute quantification relies on adequate standards. For PCR-based measurement of individual transcripts, genomic DNA can be used as a native standard, an original concatenated standard containing tandem arrays of each transcript sequence. This concept was applied to proteomics to generate peptide-concatenated standard. We are also in progress of construction of genetically modified yeast cells where unique peptide-tag adequate for mass spectrometric detection is introduced into genomic DNA encoding each protein. A strategy using unique tag has been utilized for comprehensive genetic analysis of yeast deletion library. Generation of yeast cells that have unique tags for a multitude of proteins would be feasible by use of CRISPR/Cas9 gene editing method. Asymmetrically segregation of older proteins between mother and daughter cells, possibly related to aging process, can be analyzed by combination with previously established technique for yeast cell biology. Thus, as we all know, development of unique proteomic strategies relies on a variety of classical and latest techniques and consecutively important for advance in proteomic study.