克隆编码苹果多酚氧化酶的基因组DNA并比较该基因产物在大肠杆菌和苹果中的情况。

Cloning genomic DNA encoding apple polyphenol oxidase and comparison of the gene product in Escherichia coli and in apple.

作者信息

Haruta M, Murata M, Hiraide A, Kadokura H, Yamasaki M, Sakuta M, Shimizu S, Homma S

机构信息

Department of Nutrition and Food Science, University of Tokyo, Japan.

出版信息

Biosci Biotechnol Biochem. 1998 Feb;62(2):358-62. doi: 10.1271/bbb.62.358.

DOI:10.1271/bbb.62.358

PMID:9532795

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.

摘要

克隆并测序了两个编码苹果（富士品种）多酚氧化酶（PPO）的PCR扩增基因组DNA片段。基因组DNA与cDNA的比较表明，PPO缺乏内含子。两个PPO DNA似乎都编码一个66 kDa的前体蛋白，该前体蛋白由一个56 kDa的成熟蛋白和一个10 kDa的N端转运肽组成。苹果PPO DNA在大肠杆菌中表达，通过SDS-PAGE免疫化学检测到没有转运肽的基因产物（56 kDa），其大小与苹果果实的主要PPO相同（约65 kDa）。