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KMID : 0380219930260050413
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Journal of Biochemistry and Molecular Biology
1993 Volume.26 No. 5 p.413 ~ p.419
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Expression and Purification of Fusion Proteins of Yeast ARS-Binding Factor I with Escherichia coli B-Galactosidase
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In Seop So
Guhung Jung/Hyune Mo Rho and Jiyoung Kim
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Abstract
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Yeast autonomously replicating sequence binding factor Ⅰ (ABF-1) is a multifunctional phosphoprotein which plays a role in DNA replication and transcription. We fused two different portion of yeast ABFI gene to the β-galactosidase gene and expressed the fusion proteins in E. coli cells. Two kinds of fusion proteins, ABFI(55-104)-lacZ and ABFI(55-367)-lacZ were produced under the control of E. coli tac promoter at a level of 35∼40% and 10∼15%, respectively, of total bacterial proteins. The fusion proteins retained the β-galactosidase activity. The larger fusion protein was expressed in soluble forms, while the smaller fusion protein was expressed in an 80% soluble form. The two fusion proteins were purified by β-galactosidase affinity chromatography. The fusion proteins contained the zinc finger motif derived from ABF-I protein but did not bind to the ABF Ⅰ binding site of ARS1. The results imply a possibility that if the zinc finger motif serves as DNA binding motif, other additional regions may be required for ABF Ⅰ to bind to its recognition sequence.
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