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Information Journal Paper

Title

CLONING AND EXPRESSION OF EPIDERMAL LIPOXYGENASE FROM AMBYSTOMA MEXICANUM (LOXE) IN ESCHERICHIA COLI

Pages

  2162-2170

Keywords

SODIUM DODECYL SULFATE POLYACRYLAMIDE GEL ELECTROPHORESIS (SDS-PAGE)Q2

Abstract

 Background: LIPOXYGENASE enzymes play an important role in various mechanisms of organisms. So far, many studies on human and other organisms LIPOXYGENASE activity have been conducted. In eukaryotes, this enzyme converts arachidonic acid to a variety of inflammatory mediators. For example, leukotrienes are products of this enzyme reaction. This inflammatory mediator plays an important role in the healing process. Recent studies have shown that the LIPOXYGENASE enzyme extracted from an amphibious (Ambystoma mexicanum) is more effective in the healing process in comparison with human LIPOXYGENASE. Like in the case of other enzymes, the first step for enzyme identification and characterization is to produce a large amount of purified enzyme, but the recombinant production of these proteins in bacterial expression system has not yet been reported. Therefore, in the present study we have cloned and expressed LIPOXYGENASE axolotls (LOXe) in Escherichia coli (E. coli) BL21.Methods: The sequence encoding LOXe was designed based on the amino acid sequence of the protein and then, codon optimized in order to ensure the maximum expression in E. coli. At the next step, the synthetic DNA encoding LOXe inserted into the pUC57 vector using appropriate restriction sites and then, subcloned in the PET21-A, an expression vector in order to high production of the protein in bacteria. Recombinant vector transformed to E. coli BL21 as an expression host and expression of 71kDa protein LOXe (623 amino acids) was induced in the presence of IPTG (Isopropyl-beta-D-thiogalactopyranoside).Findings: The cloning of LOXe was performed successfully and possibility of expression of this enzyme in E. coli was confirmed.Conclusion: Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicated that LOXe protein over-expressed successfully in E. coli cytoplasm.

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    APA: Copy

    TAHAN, MARYAM, MOVAHEDIAN ATTAR, AHMAD, ABBASIAN, MAHDI, & MOFID, MOHAMMAD REZA. (2014). CLONING AND EXPRESSION OF EPIDERMAL LIPOXYGENASE FROM AMBYSTOMA MEXICANUM (LOXE) IN ESCHERICHIA COLI. JOURNAL OF ISFAHAN MEDICAL SCHOOL (I.U.M.S), 31(267), 2162-2170. SID. https://sid.ir/paper/50335/en

    Vancouver: Copy

    TAHAN MARYAM, MOVAHEDIAN ATTAR AHMAD, ABBASIAN MAHDI, MOFID MOHAMMAD REZA. CLONING AND EXPRESSION OF EPIDERMAL LIPOXYGENASE FROM AMBYSTOMA MEXICANUM (LOXE) IN ESCHERICHIA COLI. JOURNAL OF ISFAHAN MEDICAL SCHOOL (I.U.M.S)[Internet]. 2014;31(267):2162-2170. Available from: https://sid.ir/paper/50335/en

    IEEE: Copy

    MARYAM TAHAN, AHMAD MOVAHEDIAN ATTAR, MAHDI ABBASIAN, and MOHAMMAD REZA MOFID, “CLONING AND EXPRESSION OF EPIDERMAL LIPOXYGENASE FROM AMBYSTOMA MEXICANUM (LOXE) IN ESCHERICHIA COLI,” JOURNAL OF ISFAHAN MEDICAL SCHOOL (I.U.M.S), vol. 31, no. 267, pp. 2162–2170, 2014, [Online]. Available: https://sid.ir/paper/50335/en

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