KMID : 1094719970020020086
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Biotechnology and Bioprocess Engineering 1997 Volume.2 No. 2 p.86 ~ p.89
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Expression of recombinant epidermal growth factor inE. coli
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Yoon Chang-Shin
Lee Eun-Gyu Lee Young-Seek Chung Il-Yup
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Abstract
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Epidermal growth factor (EGF) known as a urgastrone is a powerful mitogen with a wide variety of possibilities for medical usages. A mature EGF coding region was isolated from human prepro-EGF sequence by a conventional PCR and cloned into pQE vector in which the gene product was supposed to be expressed with 6¡¿His tag for the subsequent purification. The recombinant mature EGF was expressed in M15[Rep4], anEscherichia coli host strain, in amount of 30?40% of total proteins present inE. coli extract by the addition of isopropylthio-¥â-galactopyranoside (IPTG). The recombinant EGF purified using a Ni2+-NTA affinity column chromatography was active in its ability to induce phosphorylation on tyrosine residues of several substrate proteins when murine NIH3T3 and human MRC-5 fibroblast cells were stimulated with it. This work may provide the basic technology and information for the production of recombinant EGF.
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KEYWORD
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recombinant EGF, pQE, tyrosinephosphorylation
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