Abstract
The open reading frame encoding interferon alpha (IFN alpha) of the camel liver, Camelus dromedarius was isolated and cloned using reverse transcription-PCR. Sequence analysis of that gene showed a 564 bp encoding a protein of 187 amino acids with a predicted molecular weight of 21 kDa. Basic local alignment search tool (BLAST) sequence analysis revealed that C. dromedarius IFN alpha gene shares high sequence identity with IFN alpha genes of other species, including C. ferus, Vicugna pacos, and Homo sapiens. Expression of C. dromedarius IFN alpha cDNA in Escherichia coli revealed a fusion protein with a weight of 22.5 kDa after induction of expression with IPTG for 5 h. The recombinant IFN alpha was expressed in the form of inclusion bodies that were separated and solubilized in vitro and the protein was refolded using SDS and KCl. The folded protein is then purified using on Ni-NTA Agarose affinity chromatography and the purity was judged by SDS-PAGE. Moreover, the effect of the recombinant IFN alpha of the viability of cancer cell line was assessed by MTT assay. Morphological study showed that C. dromedarius IFN alpha protein inhibited cell survival of MDA-MB-231 triple negative breast cancer cells.