Cloning and Expression of HE4 Gene from Human Ovarian Carcinomas

Objective In order to detect the early ovarian carcinomas,we cloned the HE4 gene and constructed its expression vector.Methods The HE4 gene was amplified and cloned by RT-PCR and T-A clone techniques.The amplified HE4 gene was subcloned into expression vector PGEX-4T-1.Recombinant plasmids were identified by restriction endonuclease digestion and sequencing.The recombinant HE4 protein was expressed in TOP10 E.coli under the IPTG induction.Expressed protein was identified with Western blot analysis.Results A specific band of about 400 bp was amplified using RT-PCR from total RNA of ovarian carcinomas cells and confirmed as the HE4 gene via T-A clone and DNA sequencing analyses.SDS-PAGE showed a clear protein band with a molecular weight of 39 kDa in the IPTG-induced samples.The protein band was confirmed as HE4 protein by Western blot analysis.Conclusion The HE4 gene was successfully cloned from ovarian carcinomas cells and the HE4 protein was expressed in prokaryotic expression vector.