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

doi:10.1271/bbb.62.358

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

Research output: Contribution to journal › Article › peer-review

34 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 language	English
Pages (from-to)	358-362
Number of pages	5
Journal	Bioscience, Biotechnology and Biochemistry
Volume	62
Issue number	2
DOIs	https://doi.org/10.1271/bbb.62.358
Publication status	Published - 1998
Externally published	Yes

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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10.1271/bbb.62.358

Cite this

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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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AU - Haruta, Miyoshi

AU - Murata, Masatsune

AU - Hiraide, Ayami

AU - Kadokura, Hiroshi

AU - Yamasaki, Makari

AU - Sakuta, Masaaki

AU - Shimizu, Seki

PY - 1998

Y1 - 1998

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AB - 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.

KW - Apple (Malus pumila)

KW - Cloning genomic DNA

KW - Enzymatic browning

KW - Expression in E. coli

KW - Polyphenol oxidase

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

Abstract

Keywords

ASJC Scopus subject areas

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