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- Expression and Characterization of the Human rpS3 in a Methylotrophic Yeast Pichia pastoris
- Jae Yung Lee 1, Sang Oun Jung 2, BuHyun Youn , Oh Sik Kwon 3, Joon Kim
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Journal of Microbiology 2000;38(2):88-92.
Graduate School of Biotechnology, Korea University, Seoul 136-701, KOREA; 1 Department of Biology, College of Natural Sciences, Mokpo National Univers
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Corresponding author:
Joon Kim , Tel: 82-2-3290-3442,
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Abstract
A human ribosomal protein S3 (rpS3), which also functions as a DNA repair enzyme (UV endonuclease III), was expressed in a methylotrophic yeast, Pichia pastoris, and biochemically characterized. UV endonuclease activity was previously characterized, and this activity of mammalian rpS3 was found to be non-specific upon purification and storage. Under the Pichia expression system, the subcloned cDNA of the human rpS3 gene revealed a peptide of 42 kDa by SDS-PAGE and Western blot. The secreted form of human rpS3 rendered no endonuclease activity while the intracellular form showed UV specific endonuclease activity by the nick circle assay.
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