Production of leukemia inhibitory factor in fscherichia coli by a novel procedure and iis use in maintaining embryonic stem cells in culture

Abstract

Leukemia inhibitory factor [LIF] is a glycoprotein that is able to induce the differentiation of Ml myeloid leukemic cells and prevent the differentiation of murine embryonic stem [ES] cells. This report presents a simple method for the production and purification of large amounts of recombinant murine and human LIF in Escherichia coli. LIF is expressed initially as a fusion product with glutathione S- transferase, separated by a thrombin cleavage site. The fusion product can be rapidly purified on a glutathione-agarose affinity matrix and fully biologically active LIF released from the matrix by cleavage with thrombin. In this form LIF is suitable for many in vitro cell culture uses and in ..