Expression and Characterization of the Human rpS3 in a Methylotrophic Yeast Pichia pastoris

  • Kim, Joon (Graduate School of Biotechnology, Korea University, Seoul 136-701, KOREA) ;
  • Lee, Jae-Yung (Department of Biology, Colloge of Natural Sciences, Mokpo National University, Muan, Chonnan 534-729, KOREA) ;
  • Jung, Sang-Oun (Department of Biology, Korea University, Seoul 136-701 and Korea Institute of Science & Technology, Seoul, 136-791, KOREA) ;
  • Youn, Bu-Hyun (Department of Microbiology, College of Natural Sciences, Keimyung University, Teagu 704-701, KOREA) ;
  • Kwon, Oh-Sik (Department of Microbiology, College of Natural Sciences, Keimyung University, Teagu 704-701, KOREA)
  • Published : 2000.03.01

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 preiously characterized, and this activity of mammalian rpS3 was found to be non-specfic 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.

Keywords

References

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