Expression and characterization of recombinant single chain beta-alpha equine chorionic gonadotropin in prokaryotic host

Abstract

The aim of present study was to produce recombinant eCG/pituitary glycoprotein in the Escherichia coli (E. coli) BL21C host cells and to test its diagnostic efficacy. This aim was achieved by optimizing its expression, purification as well as its characterization through the immunoassays and bioassays. A bacterial protein expression vector system based on the phage T7 promoter and histidine tag was used for the expression and purification. The recombinant single chain beta-alpha equine chorionic gonadotropin (rbaeCG) encoding gene was constructed with beta and alpha sequences according to its biologically active counterpart. It was successfully cloned and when expressed in E.coli BL21C host, the purified recombinant protein was found to be active as revealed by enzyme linked immunosorbent assay (ELISA) and Western blotting. However, it was not found to exhibit any significant activity in vivo when tested in the mice

Molecular cloning and expression kinetics of serum interferon stimulated gene for early pregnancy detection

“…It has now become the most popular expression platform for different proteins. There are many molecular tools, protocols, a vast catalog of expression plasmids, many engineered strains and cultivation strategies which are easily available for the high-level production of heterologous proteins in E.coli (Rosano and Ceccarelli, 2014;Bhardwaj et al, 2018). Positive clones of ISG15 were introduced in expression host E. coli BL21 star (DE3) strain.…”

Characterization of the Luteinizing Hormone Beta (LH-β) Subunit Gene in the Goat

“…A recombinant single chain beta alpha equine chorionic gonadotropin/luteinizing hormone (eCG/LH) with active biological potential was produced for applications in assisted reproductive technologies or diagnostic assays through expression of the eCG/LH gene construct (Bhardwaj et al, 2018). Based on the sequence of gonadotropin subunit of goat, recombinant lutropin can be produced in an in-vitro system for long term treatment in assisted reproduction of goat without possible risk of antibody production.…”