Composition for synthesizing protein with reduced lipopolysaccharide contamination, method for producing protein using said composition

摘要

According to the present invention, a composition possessing cell-free protein synthesis activity with reduced contaminating lipopolysaccharide, and a method for producing a protein using the same are provided. When ribosome display is performed using the composition and method for protein production of the present invention, the background that is caused by non-specific binding is reduced, so that a nucleic acid that encodes the desired polypeptide can be selected with high accuracy and high efficiency.

主权项

1. A composition that possesses cell-free protein synthesis activity and that comprises an independently purified initiation factor, an independently purified elongation factor, an independently purified aminoacyl-tRNA synthetase, an independently purified ribosome, an independently purified amino acid, an independently purified nucleoside triphosphate, and an independently purified tRNA,
wherein the initiation factor, the elongation factor, the aminoacyl-tRNA synthetase, and the ribosome each were purified from a Gram-negative bacteria, wherein the composition has a ribosome concentration of 0.01- 50 μM, wherein the independently purified ribosome is a ribosome with a reduced lipopolysaccharide content, obtained by a washing step using a buffer solution comprising a surfactant, wherein the independently purified ribosome has a lipopolysaccharide concentration of 0.5 EU per pmol of ribosome or less, and wherein the composition does not comprise any release factors.