Method for producing linear DNA fragments for the in vitro expression of proteins

摘要

The present invention concerns a method for producing linear DNA fragments for the in vitro expression of proteins in which a linear DNA fragment which contains control elements necessary for expression and the protein-coding gene which has complementary regions to the two ends of the linear DNA fragment at both ends, are amplified together using a primer pair which binds upstream and downstream of the expression control regions on the linear DNA fragment, characterized in that the linear DNA fragment can be produced by linearizing an expression vector. Furthermore a method is disclosed which concerns the preparation of the protein-coding gene the ends of which have regions which overlap the linear DNA fragment. The present invention also encompasses the linear DNA fragments according to the invention for the in vitro expression of proteins, the use of these fragments, kits containing the linear DNA fragments according to the invention for the in vitro expression of proteins and methods for the in vitro expression of proteins starting from the linear DNA fragments according to the invention.