An easy cell-free protein synthesis system dependent on the addition of crude Escherichia coli tRNA
Kanda, T.; Takai, K.; Yokoyama, S.; Takaku, H.
Journal of Biochemistry 127(1): 37-41
ISSN/ISBN: 0021-924X PMID: 10731664 DOI: 10.1093/oxfordjournals.jbchem.a022581
The protein-synthesizing S30 extract of Escherichia coli contains tRNA, which limits its applications in cell-free protein synthesis. Here, we show that at least Arg- and Ser-acceptor activities can be removed from a standard S30 extract by treatment with an immobilized RNase A resin. This RNase-treated extract exhibits no protein synthesis activity, but regains it when supplied with crude E. coli tRNA and a small amount of human placental RNase inhibitor. The protein synthesis is dependent on the addition of tRNA in the presence of the RNase inhibitor. Chloramphenicol acetyltransferase was synthesized with this system and found to be active.