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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

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Publications

Construction of pPIC9 Recombinant Vector Containing Human Stem Cell Factor.

Farhadi Behrooz B   Shekari Khaniani Mahmoud M   Shekari Khaniani Mahmoud M   Mansoori Derakhshan Sima S  

Advanced pharmaceutical bulletin 20130820 2


<h4>Purpose</h4>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 h  ...[more]

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