Cloning, expression, and spectral analysis of mouse betatrophin

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GHOLAMI SAMANEH; Goodarzvand Chegini Koroush; GHEIBI NEMATOLLAH; Mokhtarian Kobra; MOHAMADI MOHSEN; FALAK REZA

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Journal: Medical Journal of The Islamic Republic of Iran Year:2017 | Volume:31 | Issue:1 Page(s): 0-0

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Title

Cloning, expression, and spectral analysis of mouse betatrophin

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Keywords

Mouse betatrophin

Recombinant protein

Refolding

CD spectroscopy

Chromatography

Abstract

Background: Betatrophin, a novel secretory protein from liver and fatty tissues, is believed to be involved in lipid and glucose metabolism. However, its precise physiological role remains unclear. Here, we report the cloning, expression, and purification steps of Mouse betatrophin in a prokaryotic system, followed by its structural analysis. Methods: Specific cloning primers were used to amplify the coding sequence of mouse liver betatrophin. The product was cloned into pET28 and expressed in E. coli BL21 (DE3) cells. The suitability of the Refolding procedure was assessed by determining secondary structures of the initial and refolded proteins using circular dichroism spectroscopy. Results: The polymerase chain reaction resulted in a 549 bp nucleotide sequence, encoding a 183 amino acid polypeptide, with an apparent molecular weight of 21 kDa, which was expressed in an inclusion body. Following an optimization and Refolding procedure, the Recombinant protein was purified by anion exchange and metal affinity Chromatography. CD spectra revealed that the refolded protein has suitable configuration. Conclusion: We believe that the produced betatrophin is suitable for further biochemical studies on glucose and lipid metabolism.

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APA: Copy

GHOLAMI, SAMANEH, Goodarzvand Chegini, Koroush, GHEIBI, NEMATOLLAH, Mokhtarian, Kobra, MOHAMADI, MOHSEN, & FALAK, REZA. (2017). Cloning, expression, and spectral analysis of mouse betatrophin. MEDICAL JOURNAL OF THE ISLAMIC REPUBLIC OF IRAN (MJIRI), 31(1), 0-0. SID. https://sid.ir/paper/294890/en

Vancouver: Copy

GHOLAMI SAMANEH, Goodarzvand Chegini Koroush, GHEIBI NEMATOLLAH, Mokhtarian Kobra, MOHAMADI MOHSEN, FALAK REZA. Cloning, expression, and spectral analysis of mouse betatrophin. MEDICAL JOURNAL OF THE ISLAMIC REPUBLIC OF IRAN (MJIRI)[Internet]. 2017;31(1):0-0. Available from: https://sid.ir/paper/294890/en

IEEE: Copy

SAMANEH GHOLAMI, Koroush Goodarzvand Chegini, NEMATOLLAH GHEIBI, Kobra Mokhtarian, MOHSEN MOHAMADI, and REZA FALAK, “Cloning, expression, and spectral analysis of mouse betatrophin,” MEDICAL JOURNAL OF THE ISLAMIC REPUBLIC OF IRAN (MJIRI), vol. 31, no. 1, pp. 0–0, 2017, [Online]. Available: https://sid.ir/paper/294890/en

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