الفهرس 
AbstractOnly 14 pages are availabe for public view
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Abstract
A 438 bp cDNA transcript coding mature ovine leptin was amplified from total adipocyte RNA by RT -PCR and cloned into the prokaryotic vector pCAL-n-EK. The recombinant vector was then transformed into XL-1 Blue E. Coli, selected and identified. Sequencing of the cloned gene showed a complete identity to the published ovine leptin. The confirmed construct was transformed into BL21 (OE3) E. Coli where protein expression was induced with IPTG. SDS-PAGE and Western blot of transformed cell lysate revealed a significant quantities of immunoreactive fusion leptin mainly in its dimeric form. Up to 6 mg/I solublely expressed leptin with more than 90% purity was purified on calmodulin-sepharose while up to 22 mg/I fusion leptin was recovered from the solubilized inclusion bodies. Both fractions were confirmed by Western blot and specific leptin radioimmunoassy.