CLONING AND EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR RECEPTOR-1 IN ESCHERICHIA COLI AND ANALYSIS OF ITS GROWTH INHIBITION EFFECT ON HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS

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AHMADVAND D.; RAHBARIZADEH F.; GERAMIZADEH B.; AHMADVAND R.; KARIMI M.H.; AKRAMI HASAN; EBADI PADIDEH

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Journal: Pathobiology Research Year:2009 | Volume:12 | Issue:1 Page(s): 1-9

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Title

CLONING AND EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR RECEPTOR-1 IN ESCHERICHIA COLI AND ANALYSIS OF ITS GROWTH INHIBITION EFFECT ON HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS

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Keywords

VEGFR-1Q3

ANGIOGENESIS INHIBITIONQ3

HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS (HUVEC)Q2

Abstract

Objective: Angiogenesis a process that results in neo-vascularization is an essential stage in growth of solid tumors and the formation of metastases. Vascular endothelial growth factor (VEGF) and its receptors, VEGFR1 (Flt-1) and VEGFR2 (KDR/Flk-1), are the major regulators for tumor angiogenesis. Recent studies showed that second domain of VEGFR-1is a key factor for VEGF/VEGFR-1 interaction. Materials and Methods: In this study, after RNA purification and cDNA synthesis, the second domain of VEGFR-1 (VEGFR-1-II) was amplified by PCR and cloned in T/A cloning vector. In order to increase the expression of the protein, we sub-cloned the gene into pET22b(+) and transformed the construct in Rosseta-gami 2, an efficient host for expression. The expression was induced by IPTG and confirmed by SDS-PAGE and Western Blotting. The recombinant protein was purified by IMAC column and the growth inhibition of human umbilical vein endothelial cells (HUVEC) was analyzed by the recombinant protein. Result: The results of SDS-PAGE and Blotting confirmed the protein purification accuracy. The recombinant protein concentration was determined by Bradford protocol. The results showed that nearly 300 mg/L VEGFR-1-II protein was produced. The function of this protein was confirmed by inhibition of HUVEC cells growth. Conclusion: Since this protein inhibited the angiogenesis in vitro it may be consider as an efficient anti-angiogenesis factor.

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APA: Copy

AHMADVAND, D., RAHBARIZADEH, F., GERAMIZADEH, B., AHMADVAND, R., KARIMI, M.H., AKRAMI, HASAN, & EBADI, PADIDEH. (2009). CLONING AND EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR RECEPTOR-1 IN ESCHERICHIA COLI AND ANALYSIS OF ITS GROWTH INHIBITION EFFECT ON HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS. PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES), 12(1), 1-9. SID. https://sid.ir/paper/80967/en

Vancouver: Copy

AHMADVAND D., RAHBARIZADEH F., GERAMIZADEH B., AHMADVAND R., KARIMI M.H., AKRAMI HASAN, EBADI PADIDEH. CLONING AND EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR RECEPTOR-1 IN ESCHERICHIA COLI AND ANALYSIS OF ITS GROWTH INHIBITION EFFECT ON HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS. PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES)[Internet]. 2009;12(1):1-9. Available from: https://sid.ir/paper/80967/en

IEEE: Copy

D. AHMADVAND, F. RAHBARIZADEH, B. GERAMIZADEH, R. AHMADVAND, M.H. KARIMI, HASAN AKRAMI, and PADIDEH EBADI, “CLONING AND EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR RECEPTOR-1 IN ESCHERICHIA COLI AND ANALYSIS OF ITS GROWTH INHIBITION EFFECT ON HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS,” PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES), vol. 12, no. 1, pp. 1–9, 2009, [Online]. Available: https://sid.ir/paper/80967/en

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