期刊
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
卷 70, 期 8, 页码 1882-1889出版社
OXFORD UNIV PRESS
DOI: 10.1271/bbb.60052
关键词
Aspergillus oryzae; EGFP; expression plasmid; protein expression; mitochondria
We report here a development of the MultiSite Gateway (TM)-based versatile plasmid construction system applicable for, the rapid and efficient preparation of Aspergillus oryzae expression plasmids. This system allows the simultaneous connection of the three DNA fragments inserted in entry clones along with a destination vector in a defined order and orientation. We prepared a variety of entry clones and destination vectors containing promoters, genes encoding carrier-proteins and fusion tags, and selectable markers, which makes it possible to generate 80 expression plasmids for each target protein. Using this system, plasmids for expression of the EGFP fused with the mitochondrial-targeting signal of citrate synthase (AoCit1) were generated. Tubular structures of mitochondria were visualized in the transformants expressing the AoCit1-EGFP fusion protein. This plasmid construction system allows us to prepare a large number of expression plasmids without laborious DNA manipulations, which would facilitate molecular biological studies on A. oryzae.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据