Patent ID: 6399323
Filing Date: 2002-06-04
Classification: C07K,C12N,C12P

Abstract:
A cell-free method of synthesizing a desired polypeptide, the method comprising:(a) adding, to a vessel comprising an ultrafiltration membrane barrier, a mixture comprising: a eukarotic cell extract capable of supporting in vitro translation, an exogenous prokaryotic RNA-polymerase, ATP, GTP, CTP, UTP, amino acids, and a DNA molecule comprising a gene encoding the desired polypeptide under the control of a promoter specific to said exogenous RNA polymerase; (b) continuously adding to the vessel the substrates ATP, GTP, CTP, UTP and amino acids, at a rate that maintains their initial concentration in the vessel; and (c) continuously removing from the vessel, through the ultra-filtration barrier, the products of the process, including AMP, GDP, CDP, UDP, pyrophosphate, inorganic phosphate, and the desired polypeptide.