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KMID : 0578319940040040467
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Molecules and Cells
1994 Volume.4 No. 4 p.467 ~ p.472
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Production and Purification of Recombinant Human Insulin-like Growth Factor I from Escherichia coli
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Kim, Sun-Ok
Park, heung-Dong/Lee, young Ik
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Abstract
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A cDNA encoding human insulin-like growth factor I (IGF-I) was cloned and expressed in Escherichia colt. IGF-I was produced as a fusion protein with the 288 N-terminal residues of P-galactosidase by a linker encoding hydroxylamine cleavage recognition sequence. The fusion protein was expressed with isopropyl-l-thio-beta-D-galactoside (IPTG) induction under the control of the inducible tac promotor in the form of insoluble inclusion bodies. After cleavage of the fusion protein with hydroxylamine, the released IGF-I was purified by a cation exchange chromatography, covalent chromatography, and reverse-phase high performance liquid chromatography (rp-HPLC). The identity and purity of the purified IGF-I was confirmed by N-terminal sequence analysis, SDS-polyacrylamide gel electrophoresis, and rp-HPLC. The biological activity of the purified IGF-I has been demonstrated to be indistinguishable from the native IGF-I in thymidine uptake, protein synthesis and radioreceptor assay.
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KEYWORD
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