Cloning genomic dna encoding apple polyphenol oxidase and comparison of the gene product in escherichia coli and in apple

Miyoshi Haruta, Masatsune Murata, Ayami Hiraide, Hiroshi Kadokura, Makari Yamasaki, Masaaki Sakuta, Seki Shimizu, Hiroshi Kadokura

Research output: Contribution to journalArticlepeer-review

31 Citations (Scopus)

Abstract

Two PCR-amplified genomic DNA fragments encoding apple (cv. Fuji) polyphenol oxidase (PPO) were cloned and sequenced. A comparison of genomic DNA with cDNAs revealed that the PPOs lacked introns. Both PPO DNAs appear to encode a 66-kDa precursor protein consisting of a 56-kDa mature protein and a N-terminal transit peptide of 10-kDa N-terminal transit peptide. Apple PPO DNA was expressed in Escherichia coli, and the gene product (56 kDa) without a transit peptide was immunochemically detected and was the same size (ca. 65 kDa) as the main PPO of apple fruit by SDS-PAGE.

Original languageEnglish
Pages (from-to)358-362
Number of pages5
JournalBioscience, Biotechnology and Biochemistry
Volume62
Issue number2
DOIs
Publication statusPublished - 1998

Keywords

  • Apple (Malus pumila)
  • Cloning genomic DNA
  • Enzymatic browning
  • Expression in E. coli
  • Polyphenol oxidase

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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