Mon. Dec 9th, 2019

ROCHE_Rapid Translation System GroE Supplement

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ROCHE

ROCHE

The Rapid Translation System (RTS) is a flexible and scalable tool for cell-free protein expression. It can be considered as “open” system, meaning that the reac-tion conditions can easily be adapted in a protein-specific manner by adding chemicals or protein factors to the reaction mixture. A very important example in this regard is the addition of chaperones in order to increase the amount of cor-rectly folded and/or soluble product.GroEL and GroES (the combination is named GroE) are cytoplasmatic chaperonins of E. coli (1,2). It is believed that they are involved in the de novo folding of 10 – 30% of all cytoplasmatic proteins (3). GroEL (57 kDa, monomer) is arranged in 2 heptameric rings which in turn form a cylindric complex. GroES (10.5 kDa, monomer) forms dome-shaped heptameric rings which cap the GroEL cylinder on one or both ends. The size of the cylinder allows proteins with a molecular mass up to 60 kDa to be substrates for the GroE sys-tem. Although it is not predictable that a particular protein is a substrate for GroE, the presence of GroE during cellular or cell-free protein synthesis has been reported in many cases to lead to higher amounts of correctly folded and active product.In order to efficiently optimize the Rapid Translation System reaction conditions and to study the effect of chaperonin addition, we generally recommend to first use the RTS 100 E.coli HY Kit (Cat. No. 3 186 148). Once a positive effect is observed in the 50 l reaction volume of RTS 100, the same conditions can be applied to the 1 ml reaction volume of the RTS 500 E.coli HY Kit (Cat. No. 3 246 817 and 3 246 949) or the RTS 500 E.coli Circular Template Kit (Cat. No. 3 018 008)

ROCHE_RTS_GroE_Supplement

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