Controlled Expression of Human Basic Fibroblast Growth Factor Mutein CS23 in Escherichia coli under a Bacteriophage T7 Promoter

概要

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A DNA sequence coding for human basic fibroblast growth factor (hbFGF) mutein CS23 was inserted down-stream of a T7 promoter in pBR322. The plasmid was introduced into Escherichia coli MM294 lysogenized with a becteriophage λhaving the T7 RNA polymerase gene under the control of the lacUV5 promoter. When 0.4 mM IPTG was added to induce gene expression in this system, the accumulation of hbFGF mutein CS23 was very rapid at the beginning, but it quickly stopped. Therefore, the total amount was low, and most of the product was inactive due to the formation of inclusion bodies. By controling the expression of the T7 RNA polymerase at an appropriate level, we succeeded in accumurating a large amount of the mutein in the cells all in a soluble and active form. By adding 0.04 mM IPTG, more than 1.5 g per liter of hbFGF mutein CS23 was accumulated under optimum culture conditions.
社団法人日本生物工学会の論文
1992-08-25