2013 Fiscal Year Final Research Report
Development of novel protein production system using peptide-tag derived from Bacillus thuringiensis Cry toxin.
| Project/Area Number |
23658048
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Applied entomology
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| Research Institution | Okayama University |
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Principal Investigator |
HAYAKAWA Tohru 岡山大学, 自然科学研究科, 助教 (30313555)
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| Project Period (FY) |
2011 – 2013
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| Keywords | Bacillus thuringiensis / Cry4Aa / 大腸菌 / ペプチドタグ4AaCter / シスタチンC / サソリトキシン |
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| Research Abstract |
4AaCter and 4AaB7 are peptide-tags derived from mosquitocidal Cry4Aa toxin. These peptide-tags facilitate inclusion formation of fused protein in Escherichia coli. In this study, the ability of 4AaCter and 4AaB7 to from protein inclusion were analyzed. Mutational analyses revealed that the hydrophobic amino acids in 4AaB7 significantly affected the formation of protein inclusion, suggesting some important roles of these hydrophobic amino acid residues. Application of 4AaCter-tag to the production of cystatin C as an endogenous marker of renal dysfunction yielded excellent results, including a dramatic increase in the production level, simplification of the product purification and high immunogenicity. Similarly, scorpion toxins were produced well as alkali-soluble protein inclusion in E. coli, and shown to be a good supplements for Cry toxin-based bioinsecticides.
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