An efficient cell-free protein synthesis system using periplasmic phosphatase-removed S30 extract

An efficient cell-free translation system was developed by removal of phosp hatase localized in the periplasmic space, which hampers the translation re action by hydrolyzing ATP. S30 extract was prepared from the spheroplast of Escherichia coli, and as much as 40% of ATP-hydrolysis activity of phospha tases could be easily removed by the spheroplast formation. The reaction pe riod of translation using phosphatase-removed S30 extract could be prolonge d and protein synthesis was enhanced by more than 30%. (C) 2000 Elsevier Sc ience B.V. All rights reserved.