Adv Pharm Bull. 2013;3(2):303-8. doi: 10.5681/apb.2013.049. Epub 2013 Aug 20.

Construction of pPIC9 Recombinant Vector Containing Human Stem Cell Factor.

Advanced pharmaceutical bulletin

Behrooz Farhadi, Mahmoud Shekari Khaniani, Sima Mansoori Derakhshan

PMID: 24312852 PMCID: PMC3848222 DOI: 10.5681/apb.2013.049

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.

Keywords: Cloning; Expression; Human SCF; Pichia pastoris

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