| Project/Area Number |
18K19187
|
|---|
| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
|
| Allocation Type | Multi-year Fund |
|---|
| Review Section |
Medium-sized Section 38:Agricultural chemistry and related fields
|
|---|
| Research Institution | Meijo University |
|---|
Principal Investigator |
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
加藤 晃代 名古屋大学, 生命農学研究科, 招へい教員 (40727640)
|
|---|
| Project Period (FY) |
2018-06-29 – 2020-03-31
|
| Project Status |
Completed (Fiscal Year 2019)
|
| Budget Amount *help |
¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2019: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2018: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
|
|---|
| Keywords | リボソームタンパク質 / 発現量 / タンパク質生産 / タンパク質発現系 / 異種発現 |
|---|
| Outline of Final Research Achievements |
There are no general rule about which is the best platform for each protein of interest. Therefore ‘How we can regulate protein expression at any expression platform?’ is the one of the biggest issues that no one can answer.
The purpose of this research project is to find out a rule that can easily express the target protein in any expression system. In this research, we constructed original database focused on the N-terminal sequences of ribosomal subunits protein in various species.First, database of theoretical m/z of ribosomal subunits protein in a total of about 50 types of various species were constructed. Next, the whole cell fraction was subjected to MALDI-TOF MS analysis.
As a result, specific amino acid residue, Ala, Lys, Ser, Val, and Thr, in their N-terminal amino acid frequency was high. The same tendency was observed in the other strains analyzed. This suggests that the nascent amino acids may be used as an indicator of proteins that are highly produced in each species.
|
| Academic Significance and Societal Importance of the Research Achievements |
現在、どの発現系を用いれば目的タンパク質を自由自在に組換え生産できるかのルールが無く、経験と運に頼らざるをえない。
そのルールを見出すため、質量分析計で検出可能なリボソームタンパク質と、そのN末端付近のアミノ酸配列に着目し、より検出シグナルの高いものとその配列を解析した。その結果、N末端の傾向は、Ala、Lys、Ser、Val、Thrの偏りが強いことが判明し、各生物種で生産されやすいタンパク質の指標として新生鎖のアミノ酸を利用できる可能性が示唆され、目的タンパク質の発現ルールが存在する可能性を見出すことできた。本成果の発展は、自由自在な目的タンパク質の生産に貢献でき、社会的インパクトが大きい。
|
