Preparation of human EGFR intracellular domain-GST fusion protein with recombinant technique

EGFR subtypes have been found overexpressed in many GI tumours. Recently, a great number of inhibitory compounds acting on EGFR have been identified. Our aim is to screen large numbers of inhibitory compounds with an ELISA-based tyrosine kinase assay established in our laboratory. To meet the needs of high amount EGFR enzyme arising in this assay technology, custom production of the recombinant enzyme is necessary. To establish a recombinant expression system, we isolated mRNA from human A431 cells and EGFR intracellular domain was amplified with PCR reaction using primers with Not1 cleavage sequences. PCR produ…