| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2017: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Outline of Final Research Achievements |
In this research, a novel technique was developed which enabled the refolding of proteins at high temperatures with catalysis by thermostable protein disulfide isomerase. For this purpose a novel reporter protein sfGFP-2SS was invented, which emits green fluorescence depending on restoration of correct S-S bonds to be introduced. The fluorescence can be once vanished by erroneous S-S crosslink upon oxidation under denaturation, but green fluorescence was restored by disulfide isomerase from Thermus thermophilus HB8 in the programmed thermal cycling with redox buffer GSH/GSSG. The invention of Protein Thermal Refolding technique, termed as PTR, was established as a protein version of PCR, leading denatured proteins with abbrenet disulfide bonds to the correct structure with high expectation for recombinant antibody production.
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