Riken Corporation

Goushi Ishihara

Shigeyuki Yokoyama

Mikako Shirouzu

Takanori Kigawa

Kaori Tajima

Takashi Yabuki

Mie Goto

Mihoro Saeki

We claim a method of expression a thioredoxin-fused membrane protein comprising expressing said thioredoxin-fused membrane protein in a cell-free protein synthesis system in the presence of a non-ionic detergent. It is preferable that the fused membrane protein which is a highly hydrophobic protein, e.g., a G protein-coupled receptor, can be produced in a state being highly soluble and capable of forming a biologically active three-dimensional structure. Also, this invention provides a recombinant polynucleotide encoding said thioredoxin-fused membrane protein which is expressed in the cell-free protein synthesis system.

Method for producing by cell-free protein synthesis system using thioredoxin-fused protein expression vector

A method for producing a protein using a cell-free protein synthesis system comprising a detergent so that the protein can be synthesized without aggregation, is provided. The protein is a protein comprising a hydrophobic region in at least a portion thereof, for example, a membrane protein or its fragment (portion). And the detergent is a mild detergent which would not denature the protein, for example, a nonionic or amphoteric ionic detergent.

Process for synthesizing protein using cell-free protein synthesis system

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Yokohama, Japan

Goushi Ishihara