Cloning genomic DNA encoding apple polyphenol oxidase and comparison of the gene product in Escherichia coli and in apple.
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.
Haruta
M
M
Department of Nutrition and Food Science, University of Tokyo, Japan.
Murata
M
M
Hiraide
A
A
Kadokura
H
H
Yamasaki
M
M
Sakuta
M
M
Shimizu
S
S
Homma
S
S
D000595:Amino Acid Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D004156:Catechol Oxidase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D007438:Introns; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D019685:Rosales / Q000201:enzymology*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D004156:Catechol Oxidase