State analyses of a cell-free protein synthesis system using ribosome profiling

2016 Fiscal Year Final Research Report

• Project/Area Number: 26640134
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: System genome science
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Shimizu Yoshihiro 国立研究開発法人理化学研究所, 生命システム研究センター, ユニットリーダー (90401231)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 無細胞タンパク質合成 / リボソーム / mRNA / 次世代シーケンサー

Outline of Final Research Achievements

We aimed to establish a technique to analyze how the internal state of the PURE system, a reconstituted cell-free protein synthesis system, changes according to the changes in mRNA sequence by using deep sequencing. By preparing an mRNA where 12 bases downstream of the start codon are randomized and translating it in the PURE system to obtain an initiation complex or elongation complex, only the mRNA sequence interacting with the ribosome is specifically extracted. It became possible to quantitatively evaluate the influence of mRNA sequence by performing deep sequence analysis.

Free Research Field

生化学