Construction of pPIC9 Recombinant Vector Containing Human Stem Cell Factor.

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.

SUBMITTER: Farhadi B 

PROVIDER: S-EPMC3848222 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature