Differential fusion expression and purification of a cystatin in two different bacterial strains.
To date, the identification of the novel multifunctional properties of cysteine proteinase inhibitors "known as cystatins" is the great of interests for molecular biologists. The efficient production, purification and correctly folded form of these proteins are the most important requirements for their any basic research. To the best of our knowledge, maltose-binding protein (MBP) fusion tags are being used to overcome the impediment to their heterologous recombinant expression in Escherichia coli as insoluble and bio-inactive inclusion bodies. In the present work, to evaluate the expression efficiency of a cystatin molecule in E. coli cells by using MBP tags, the expression of Celosia cystatin was studied in two different strains of this bacterium. The quantitative analysis results based on the one-step purification yield of the fused product showed the excellency of the E. coli TB1 strain in comparison to E. coli DH5alpha for the high-level production of active product.
Research Institute for Fundamental Sciences (RIFS), University of Tabriz, Tabriz, Iran. aghz_bioch@yahoo.co.in
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Gholizadeh
D001483:Base Sequence; D034762:Celosia / Q000737:chemistry; D003001:Cloning, Molecular / Q000379:methods; D015891:Cystatins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry*; D058619:Maltose-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D012995:Solubility; D013045:Species Specificity
D015891:Cystatins; D018076:DNA, Complementary; D058619:Maltose-Binding Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins