Recombinant expression and purification of functional vascular endothelial growth factor-121 in the baculovirus expression system

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• 作者：Nastaran Mohseni¹ ; Ali Jahanian-Najafabadi² ; Fatemeh Kazemi-Lomedasht¹ ; Roghaye Arezomand³ ; Mahdi Habibi-Anbouhi⁴ ; Delavar Shahbazzadeh¹ ; Mahdi Behdani¹ ; ^{Behdani73042@yahoo.com} ; ^{Behdani@pasteur.ac.ir}
• 关键词：Vascular endothelial growth factor ; Baculovirus expression system ; Recombinant bacmid
• 刊名：Asian Pacific Journal of Tropical Medicine
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：9
• 期：12
• 页码：1195-1199
• 全文大小：1077 K
• 卷排序：9

文摘

To express human vascular endothelial growth factor121 (VEGF121) in insect cells.MethodsA gene construct containing VEGF was cloned in the pFastBac-HTA vector, followed by transformation in DH10BAC. The recombinant bacmid was then extracted, and transfected into Sf9 insect cells. The transfected cells were harvested, and then VEGF expression was confirmed by western blotting using specific antibodies. The tube formation assay was used for functional assessment of VEGF.ResultsOur results showed that VEGF could be successfully expressed in the baculovirus system. Purified VEGF was able to stimulate in vitro tube formation of human endothelial cells.ConclusionsResults from this study demonstrated that the recombinantly-produced VEGF can be considered as a promising candidate for therapeutic purposes.