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:: Volume 22, Issue 1 (spring2012 2012) ::
MEDICAL SCIENCES 2012, 22(1): 32-38 Back to browse issues page
Cloning and expression of placental growth factor protein of human -1 (hPLGF-1) in Rosetta E.coli expression system
Narges Arbabi1, Mehdi Behdani2, Majid Golkar3, Seyed Hamid Aghaee bakhtiari3, Hossein Khan ahmad shahreza4, Reza Mahdian *5
1- Islamic Azad University, Science and Research Campus, Tehran, Iran
2- , Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
3- , Pasteur Institute of Iran, Tehran, Iran
4- Pasteur Institute of Iran, Tehran, Iran
5- Pasteur Institute of Iran, Tehran, Iran , rezamahdian@yahoo.com
Abstract:   (16480 Views)
Background: Angiogenesis or new blood vessels generation is the most important factor affecting cell growth and proliferation in physiologic and pathologic conditions. Placenta Growth Factor (PLGF) is one of the important proteins for angiogenesis induction. In this study, placenta-derived PLGF-1 cDNA was cloned and expressed in Escherichia coli expression system.
Materials and methods: In this experimental study, the human placenta-derived PLGF-1 gene was amplified using specific primers and was cloned into pET32a and pET28a expression vectors. In order to express target protein, pET32a-PLGF-1 and pET28a-PLGF-1 constructs were transformed into E.coli Rosetta. PLGF-1 expression was induced by IPTG, and then the yield of expressed protein and its solubility was analyzed by SDS-PAGE and Western blotting assay.
Results: The pET32a-PLGF-1 and pET28a-PLGF-1 constructs were confirmed by sequencing. The bacterial lysates derived from IPTG-induced samples showed exact proteinand confirmed by western bloting. The majority of the expressed protein was insoluble. Conclusion: The PLGF-1 is well expressed in Rosetta E.coli system and it could be used in different project.
Keywords: Angiogenesis, Cloning, Gene expression, PLGF-1
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Semi-pilot: Experimental | Subject: Genetic
Received: 2012/04/3
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arbabi N, Behdani M, golkar M, Aghaee bakhtiari S H, Khan ahmad shahreza H, Mahdian R. Cloning and expression of placental growth factor protein of human -1 (hPLGF-1) in Rosetta E.coli expression system. MEDICAL SCIENCES. 2012; 22 (1) :32-38
URL: http://tmuj.iautmu.ac.ir/article-1-541-en.html


Volume 22, Issue 1 (spring2012 2012) Back to browse issues page
فصلنامه علوم پزشکی دانشگاه آزاد اسلامی واحد پزشکی تهران Medical Science Journal of Islamic Azad Univesity - Tehran Medical Branch
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