Construction of pPIC9 Recombinant Vector Containing Human Stem Cell Factor
Behrooz Farhadi1, Mahmoud Shekari Khaniani, Sima Mansoori Derakhshan
1Department of Biochemistry, Faculty of Medicine, Tabriz University of Medical Sciences Tabriz, Iran.
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.
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