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6th World Congress on Biotechnology

New Delhi, India

Anuradha Bhardwaj

Anuradha Bhardwaj

ICAR-National Research Centre on Equines, India

Title: Expression and characterization of eCG recombinant protein in Sf cell lysate

Biography

Biography: Anuradha Bhardwaj

Abstract

The family of heterodimeric glycoprotein hormones (LH, FSH, TSH, CG, Inhibins) play imperative role in reproduction. These hormones are composed of two dissimilar subunits: Alpha and beta. The alpha subunit is generally common to all glycoprotein hormones, however; the beta subunit is species specific and accountable for receptor binding specificity. In the present study, the eCG gene was synthesized with signal sequence of beta and C-terminus of alpha subunits together and amplified with polymerase chain reaction with specific primer pairs. The amplified fragment (~700 bp) was gel purified and successfully cloned in pUC57 cloning vector. The cloned construct got confirmed with sequencing and sub-cloned in pIX 4.0 vector for expression in Spodoptera frugiperda insect cell lysate. Expression was carried out as per recommended protocol of EasyXpress® Insect Cell Protein Synthesis kit. Recombinant protein of ~27 kDa was analyzed in SDS PAGE and characterized by ELISA. This study paves way for new generation of cell free expression and purification recombinant hormones for further study.