Author/Authors :
Farhadi، Behrooz نويسنده Department of Biochemistry, Faculty of Medicine, Tabriz University of Medical Sciences Tabriz, Iran. , , Shekari Khaniani، Mahmoud نويسنده MD, PhD, Assistant Professor, Department of Human Genetics, Faculty of Medicine, Tabriz University of Medical Sciences,Tabriz, Iran , , Mansoori Derakhshan، Sima نويسنده Department of Medical Genetics, Faculty of Medicine, Tabriz University of Medical Sciences Tabriz, Iran. ,
Abstract :
Purpose: Various cytokine regulates hematopoesis; they promote number of stages in stem cells biology such as proliferation, differentiation and endurance. Biological effects of SCF, as a hematopoietic cytokine; is triggered by binding to its ligand c-kit. Potential therapeutic applications of SCF include hematopoietic stem cell mobilization, exvivo stem/progenitor cell expansion, gene therapy, and immunotherapy. In this study we tried to construct of pPIC9 recombinant vector containing human SCF. Methods: hSCF cDNA was amplified by PCR and both hSCF cDNA and pPIC9 as yeast expression vector (shuttle vector) digested by EcoR I and Xho I restriction enzymes. Subsequent the digestion reaction, ligation reaction was carried out. In order to verifying of pPIC9 recombinant vector containing hSCF, PCR and sequence analysis was performed. Results: The construction of recombinant expression vector of pPIC9 containing hSCF cDNA was confirmed by sequencing method successfully. Conclusion: rhSCF/pPIC9 vector can be transformed into the Picha pastoris yeast as a eukaryotic host in order to produce human SCF at industrial scale.
