Abstract
The mature gene of gloshedobin, a snake venom thrombin-like enzyme from the snake, Gloydius shedaoensis, was cloned and expressed in strain E. coli BL21 (DE3). Having been induced by IPTG, the recombinant gloshedobin was in both soluble and insoluble forms. To avoid inclusion body formation, expression was optimized at 25 degrees C. Furthermore, a 50% increase in solubilization of the target protein was obtained by adding 0.1 mM Mg2+ to the medium. The purified recombinant gloshedobin gave a 44 kDa band on SDS-PAGE gel.
MeSH terms
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Animals
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Cells, Cultured
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Enzyme Stability
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Escherichia coli / drug effects
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Escherichia coli / enzymology*
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Escherichia coli / genetics
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Gene Expression Regulation, Bacterial / drug effects
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Gene Expression Regulation, Enzymologic / drug effects
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Ions
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Metals / pharmacology*
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Quality Control
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / chemistry
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Recombinant Proteins / classification
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Recombinant Proteins / isolation & purification
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Temperature
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Thrombin / biosynthesis*
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Thrombin / chemistry
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Thrombin / classification
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Thrombin / isolation & purification
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Viper Venoms / biosynthesis*
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Viper Venoms / chemistry
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Viper Venoms / classification
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Viper Venoms / isolation & purification
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Viperidae / metabolism
Substances
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Ions
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Metals
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Recombinant Proteins
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Viper Venoms
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Thrombin
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gloshedobin protein, Gloydius shedaoensis