Heterologous Expression and Characterization of Indian Sahiwal Cattle (Bos indicus) Alpha Inhibin

Abstract

Inhibin is a non-steroidal glycoprotein hormone of gonadal origin with major action as negative feedback control of the production of FSH by the anterior pituitary gland. The physiological role of inhibin has led to the development of inhibin immunogens for fertility enhancement in farm animals. It is envisaged that a reduction of endogenous inhibin secretion would increase FSH concentrations and thus offers a potential for increasing the number of ovulatory follicles in the ovary. The present work was carried out to produce recombinant bovine (Indian Sahiwal Cattle; Bos indicus) alpha inhibin (bINH-α) in E. coli by optimizing its expression and purification in biologically active form and to study its immunological characterization. A bacterial protein expression vector system based on the phage T(5) promoter was used. The bINH-α encoding gene was successfully cloned and expressed in E. coli and the purified recombinant bINH-α was characterized. Recombinant bINH-α (25 µg mL(-1)) immunized guinea pigs had a significant increase in litter size compared to the control group. These results indicate a role for recombinant bINH-α as a fecundity vaccine to enhance the ovulation rate and litter size in animals.

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

“…The recombinant full length single chain reCG showed good results in ELISA and Western Blot, which emphasized its diagnostic potential and affinity towards antibody binding. Earlier, the lead author had expressed and characterized the bovine inhibin alpha subunit in E.coli M15 cells (Bhardwaj et al, 2012). There too, difficulties were encountered in purifying the recombinant protein from inclusion bodies.…”

“…Seo et al, 2015, focused on the biological activities (LHand FSH-like activities) of amino acids (94, 95, and 96) in the eCG -subunit. In the present work, we carried out the expression, purification and characterization of single chain beta alpha eCG through bacterial expression system on similar lines of our earlier work on recombinant bovine inhibin alpha (Bhardwaj et al, 2012(Bhardwaj et al, , 2013. Recently, we reported the characterization and molecular docking analysis of eCG alpha molecule (Bhardwaj et al, 2017).…”

Trends in recombinant protein use in animal production

“…However, in many other cases inhibin has been produced in mammalian cells [16, 92]. Importantly, recombinant inhibin has been used as an antigen for the immunization against endogenous inhibin in hens [93, 94], cockerels [95], heifers [87, 89, 96], water buffaloes [88], guinea pigs [97], goats [98], and sheep [99–102]. Only in a minority of cases, the recombinant inhibin α subunit used is obtained synthetically [103–106] or purified from follicular fluid [107].…”