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; Seiichi HOMMA

doi:10.1271/bbb.62.358

Food & Nutrition Notes

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, Seiichi HOMMA

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Keywords: apple (Malus pumila), enzymatic browning, polyphenol oxidase, cloning genomic DNA, expression in E. coli

JOURNAL FREE ACCESS

1998 Volume 62 Issue 2 Pages 358-362

DOI https://doi.org/10.1271/bbb.62.358

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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.

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