2016 Fiscal Year Final Research Report
Definition and optimization of secretion signal by using artificial model sequences in yeast and human cells
| Project/Area Number |
26450505
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Applied molecular and cellular biology
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| Research Institution | Yamaguchi University |
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Principal Investigator |
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| Project Period (FY) |
2014-04-01 – 2017-03-31
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| Keywords | 分泌シグナル / Kluyveromyces marxianus / 網羅的変異解析 / 人工配列 / ヒト培養細胞 |
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| Outline of Final Research Achievements |
A secretory signal sequence located at the N-terminus of a protein directs the protein to the extracellular environment. Signal sequences usually contain an N-terminal basic amino acid followed by a hydrophobic stretch, although no consensus signal sequence has been identified. In this study, mutagenesis and simple modeling of signal sequences was attempted to define optimum signal sequence. Mutagenesis revealed the importance of N-terminal basic residue, length of hydrophobic stretch and Glu following to the stretch in yeast. Hydrophobic stretch region can be substituted with artificial sequence consisting of a repeat of a single hydrophobic amino acid. Stretches consisting of Phe, Leu, Ile, or Met were effective for secretion but the number of residues affected secretory activity. A stretch containing methionine residues showed the highest activity. In human cultured cell, definition of secretion signal is similar but looser compared with the yeast.
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| Free Research Field |
遺伝子工学
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