Abstract: Here we describe a new modified overlap extension PCR method for cloning human tau isoforms which can cut and splice gene to make inserts for expression vectors easily. The isoforms could be simply cloned to eukaryotic expression vector pcDNA4 and the tau protein expression be detected by western blot. Our data implicate that this method is much convenience and easier to cut and splice the multiple cloning site without restriction enzyme. It possesses great flexibility, valuablity and may provide broad applications in molecular cloning.