:: Volume 7, Number 2 (Summer 2017) ::
J Fasa Univ Med Sci 2017, 7(2): 223-232 Back to browse issues page
Codon Optimization of Stem Cell Factor (SCF) gene and evaluation of this gene expression using vector pBudCE4.1 in Chinese Hamster Ovary cells (CHO)
ehsan heidari1, maryam peymai 2, ali kazemi3, kamran Ghaedi4
1- Department of Biology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Sahrekord, Iran
2- Department of Chemistry, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Sahrekord, Iran , m.peymani62@gmail.com
3- Department of Chemistry, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Sahrekord, Iran
4- Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran
Abstract:   (171 Views)

Background & Objective: Stem cell factor (SCF) is a blood cytokine which can play a significant role in the differentiation of blood precursor cells, survival, proliferation and differentiation of mast cells, and it can also increase the proliferation and invasion of tumor cells through affecting hematopoietic cells. Its therapeutic effect in the treatment of diseases such as Alzheimer's and myocardial infarction is being investigated. The aim of this study was to clone SCF gene into the pBudCE4.1 expression vector by codon optimization in Chinese hamster ovary cells (CHO).
Materials & Methods: In this experimental study, SCF sequence was made synthetically after codon optimization for CHO cells and changing of the contents of G+C and subcloned into the expression vector pBudCE4.1. Then, SCF gene was transfected into CHO-K1 cells and evaluated using RT-qPCR for gene expression of the relevant gene.
Results: Recombinant expression vector pBudCE4.1/SCF was constructed and approved by PCR technique, enzyme digestion and sequencing. RT and PCR results showed that SCF genes can be expressed in CHO cells after codon optimization.
Conclusion: The results showed that the cloning of SCF gene can be done well through expression vector pBudCE4.1 and this expression vector can be an appropriate vector for expressing SCF gene into CHO cells.
 

Keywords: Codon optimization, Cloning, Chinese Hamster Ovary Cells (CHO), Stem Cell Factor (SCF)
Full-Text [PDF 1656 kb]   (21 Downloads)    
Type of Study: Research | Subject: Cellular-Molecular Biology
Received: 2016/11/3 | Accepted: 2017/03/9 | Published: 2017/09/4



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Volume 7, Number 2 (Summer 2017) Back to browse issues page