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Mammalian cells are the recommended host for recombinant eukaryotic protein production aimed at incorporation of post-translational modifications for downstream applications. The bacterial system and insect cells are widely used because of ease of technical methodology, economics of production, purification and yield of final protein. The present research objective was expression of recombinant reproductive hormones of animal origin and study of their immunogenic potential for reproductive applications. The equine Chorionic Gonadotropin (eCG) is one of the most heavily glycosylated protein amongst all glycoprotein hormone family. Hence, experiments were carried out to observe its expression in the three most popular host systems and it led to comparative studies for their post-translational modifications. The Pregnant Mare Serum Gonadotropin (PMSG, also called as eCG) gene was cloned in TOPO-TA vector, pIX 4.0 and pTARGET vectors accordingly and expression analysis in E. coli BL21C, Sf insect cell lysate and COS-1 cells was carried out. We observed diverse sizes of recombinant proteins in SDS-PAGE analysis which indicated post-translational modification in mammalian expression system towards the linking of tags as well as side chains in respective host cells. Basic diagnostic immunogenicity tests showed encouraging results, however, no significant in vivo and in vitro activity was observed for the expressed reCG in all the employed host systems.

To establish normal values of blood biochemical indices for different indigenous local donkey population available in various agro-climatic zones, blood samples were collected from 233 adult and apparently healthy donkeys. The samples were analysed for metabolites (albumin, total serum protein, direct bilirubin, total bilirubin, cholesterol, HDL cholesterol, urea, uric acid, triglyceride, creatinine) and minerals (calcium, phosphorus) to evaluate significant difference within and between populations. Confidence limit of each biochemical indices showed a close range as compared to their actual range observed under varied geographic areas. All the metabolites and minerals showed significant variations in their levels within and between donkey populations which could possibly be due to the differences in the nutritional status of donkeys, their managemental aspects and biochemical metabolism. In agro-climatic zone 1 (Spiti and Leh areas), having low vegetation cover with poor nutritious grasses for a limited period, levels of most of the biochemical profiles in donkey populations belonging to these areas were significantly lower than those in other zones (VI, IX, XII, XIV). This study indicated that normal biochemical values of different indices for a particular population should not be used as such for disease prognosis, diagnosis and therapeutic monitoring of other donkey population belonging to other agro-climatic zone having different nutritional and managemental practices.

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

Trypanosoma evansi (T. evansi) is a causative agent of disease called surra, affecting wide range of domestic and wild animals. In this study, a PCR assay was developed using primers targeting ITS-1 region flaking between 5.8 S and 18 S subunits of rRNA. The test was employed using serially diluted DNA extracted from purified trypanosomes ranging from 200 ng/μl to 0.00002 pg/μl and T. evansi infected mice blood ranging from 350 ng/μl to 0.00035 ng/μl. The diagnostic sensitivity of PCR assay was found to be 0.2 pg/μl and 0.035 ng/μl with purified parasite DNA and infected mice blood DNA, respectively.