Establishing an efficient gene-targeting system in an itaconic-acid producing Aspergillus terreus strain.
OBJECTIVE
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.
Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Synthetic Biology, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Qingdao, 266101, China.|University of Chinese Academy of Sciences, Beijing, 100049, China.
Xuenian
X
Huang
Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Synthetic Biology, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Qingdao, 266101, China.
Mei
M
Chen
Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Synthetic Biology, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Qingdao, 266101, China.|National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, 100190, China.
Jianjun
J
Li
Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Synthetic Biology, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Qingdao, 266101, China. lvxf@qibebt.ac.cn.
Xuefeng
X
Lu
D001230:Aspergillus / Q000378:metabolism*; D018390:Gene Targeting / Q000379:methods*; D013386:Succinates / Q000378:metabolism*
D013386:Succinates; C005229:itaconic acid
Aspergillus terreus; Cre/loxP system; Gene-targeting; Itaconic acid; Non-homologous end joining; pyrG