Paper Information

Journal:   ADVANCED PHARMACEUTICAL BULLETIN   2013 , Volume 3 , Number 2; Page(s) 303 To 308.
 
Paper: 

CONSTRUCTION OF PPIC9 RECOMBINANT VECTOR CONTAINING HUMAN STEM CELL FACTOR

 
Author(s):  FARHADI BEHROOZ, SHEKARI KHANIANI MAHMOUD, MANSOORI DERAKHSHAN SIMA*
 
* 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.

 
Keyword(s): HUMAN SCF, CLONING, EXPRESSION, PICHIA PASTORIS
 
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