Establishing an efficient gene-targeting system in an itaconic-acid producing Aspergillus terreus strain
Establishing an efficient gene-targeting system in an itaconic-acid producing Aspergillus terreus strain
复制标题
在产衣康酸土曲霉菌株中建立有效的基因靶向系统
DOI：
10.1007/s10529-016-2143-y
复制
发表时间：
2016-09-01
影响因子：
2.7
通讯作者：
Lu, Xuefeng
中科院分区：
文献类型：
--
作者：
Huang, Xuenian;Chen, Mei;Lu, Xuefeng
目的：构建一个高效的基因打靶平台，用于筛选高产衣康酸的土曲霉CICC 40205。结果：通过敲除ku 80基因，提高了同源重组的频率。在ku 80-/pyrG-双突变体中建立了一个基于双向选择标记pyrG An的营养转化系统，便于后续标记拯救。利用改良的Cre/loxP重组系统，通过将Cre重组酶直接导入原生质体，切除pyrG An标记。有利于研究生物合成机理，提高衣康酸等产品的生产能力。
Objectives:To develop an efficient gene-targeting platform in an excellent itaconic acid producing strain Aspergillus terreus CICC40205.Results:The frequency of homologous recombination was improved by deleting the ku80 gene. A nutritional transformation system based on the bidirectionally selectable marker, pyrG An , was established in the ku80-/pyrG-double mutant which is convenient for following marker rescue. The modified Cre/loxP recombination system was applied for the excision of the pyrG An marker by directly introducing Cre recombinase into the protoplasts.Conclusions:This gene-targeting system is an efficient platform for sequential and multiple genetic modifications in A. terreus and is conducive to study biosynthesis mechanisms and to improve the production ability of itaconic acid and other products.