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:: Volume 3, Issue 1 (Winter - 2015) ::
Shefaye Khatam 2015, 3(1): 21-28 Back to browse issues page
Subcloning of NGF Gene into pSecTag2/Hygro Secretory Vector and Expression in PC12 Cell Line
Bahman Jalali Kondori *, Mohammad Hossein Asadi, Fateme Azemati
Department of Anatomical Sciences, School of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran. , Bahmanjalali2010@gmail.com
Abstract:   (7780 Views)

Introduction: Subcloning of specific genes in plasmids and transfecting them into target cells for producing therapeutic proteins or cell differentiation, is suggested as one of the most effective treatment methods for different diseases. Nerve growth factor (NGF) is one of the members of the neurotrophin family. Neurotrophins are a family of proteins that regulate the survival, differentiation, and function of different types of neurons. Studies showed that NGF gene expression in stem cells induces differentiation toward neuron-like cells as well as the growth of axons and its branching. Materials and Methods: In this research, enzymatic digestion on a plasmid carrying NGF gene was performed and extracted. NGF gene was subcloned into pSecTag2/Hygro secretory plasmid. The subcloned plasmid was precipitated and concentrated. It was then transfected by Lipofectamine into PC12 cell line. NGF gene expression and protein production were evaluated using RT-PCR and western blot methods. Results: Sequence determination indicated that secretory plasmid subcloning process has been correct. Expression of NGF gene in transfected PC12 cells was shown by RT-PCR method and production of its protein was proved by the results from western blots. Conclusion: Subcloning of NGF gene in pSecTag2 secretory vector is a suitable technique for transfer to eukaryotic cells.

Keywords: Transfection, Genetic Vectors, Nerve Growth Factors, PC12 Cells
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Type of Study: Research --- Open Access, CC-BY-NC | Subject: Molecular Neurobiology

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Jalali Kondori B, Asadi M H, Azemati F. Subcloning of NGF Gene into pSecTag2/Hygro Secretory Vector and Expression in PC12 Cell Line. Shefaye Khatam. 2015; 3 (1) :21-28
URL: http://shefayekhatam.ir/article-1-645-en.html

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Volume 3, Issue 1 (Winter - 2015) Back to browse issues page
مجله علوم اعصاب شفای خاتم The Neuroscience Journal of Shefaye Khatam
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