Indian Journal of Animal Research

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Research Article

Indian Journal of Animal Research, volume 53 issue 5 (may 2019) : 587-593

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

Anuradha Bhardwaj, Sanjay Kumar, Varij Nayan, Parvati Sharma, Yash Pal, S.C. Yadav
1ICAR-National Research Centre on Equines, Sirsa Road, Hisar-125 001, Haryana, India.
Cite article:- Bhardwaj Anuradha, Kumar Sanjay, Nayan Varij, Sharma Parvati, Pal Yash, Yadav S.C. (2018). Expression and characterization of recombinant single chain beta-alpha equine chorionic gonadotropin in prokaryotic host. Indian Journal of Animal Research. 53(5): 587-593. doi: 10.18805/ijar.B-3571.

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

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