Expression and characterization of the human rps3 in a methylotrophic yeast Pichia pastoris

Jae Yung Lee, Sang Oun Jung, Bu Hyun Youn, Oh Sik Kwon, Joon Kim

Research output: Contribution to journalArticle

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.

Original languageEnglish
Pages (from-to)88-92
Number of pages5
JournalJournal of Microbiology
Volume38
Issue number2
Publication statusPublished - 2000

Keywords

  • Endonuclease activity
  • Nick circle assay
  • Pichia pastoris
  • rpS3

ASJC Scopus subject areas

  • Microbiology
  • Applied Microbiology and Biotechnology

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