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KMID : 0578319940040010125
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Molecules and Cells
1994 Volume.4 No. 1 p.125 ~ p.129
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synthesis of Active Tadpole H-chain ferritin in Escherichia coli
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Kim Young-Taek
Kim Kyung-Suk
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Abstract
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We established the method to produce large amounts of tadpole H-chain fenitin in Escherichia coll. The coding region of the cDNA for the ferritin H-chain was prepared by oligodeoxynucleotide-directed deletion and inserted into the plasmid pKK223-3 and further transferred to the multi-copy plasmid pVUCH-1, producing the high-level expression vector pVUTFHIO. The expression of the ferritin H-chain gene was examined by SDS-PAGE, which resulted in a band at 22 kDa. Although both plasmids pKKTFHIO and pVUTFHIO contain the same tac promoter, the expression yields were different. Using the latter vector, the proteins, expressed as approximately >30% of the soluble proteins, were assembled into complete ferritin and were able to uptake iron. This band was positively confirmed by Western blotting, proving that the expressed protein shares an antigenic determinant with native tadpole ferritin.
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