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Expression and characterization of eCG recombinant protein in Spodoptera frugiperda cell lysate
6th World Congress on Biotechnology
Anuradha Bhardwaj, Sanjay Kumar and A K Gupta
ICAR-National Research Centre on Equines, India
ScientificTracks Abstracts: J Biotechnol Biomater
DOI:
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�?庐 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.Biography
Anuradha Bhardwaj has completed her PhD from ICAR-NDRI in Animal Biotechnology Discipline and she is currently working as a Scientist (Biotechnology-Animal Sciences) at National Research Centre on Equines (ICAR), India.
Email: dranu.biotech@gmail.com
