title: A Tet-on and Cre-loxP Based Genetic Engineering System for Convenient Recycling of Selection Markers in Penicillium oxalicum.
abstract: The lack of selective markers has been a key problem preventing multistep genetic engineering in filamentous fungi, particularly for industrial species such as the lignocellulose degrading Penicillium oxalicum JUA10-1(formerly named as Penicillium decumbens). To resolve this problem, we constructed a genetic manipulation system taking advantage of two established genetic systems: the Cre-loxP system and Tet-on system in P. oxalicum JUA10-1. This system is efficient and convenient. The expression of Cre recombinase was activated by doxycycline since it was controlled by Tet-on system. Using this system, two genes, ligD and bglI, were sequentially disrupted by loxP flanked ptrA. The successful application of this procedure will provide a useful tool for genetic engineering in filamentous fungi. This system will also play an important role in improving the productivity of interesting products and minimizing by-product when fermented by filamentous fungi.
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University Jinan, China.
authors.forename: Baojie
authors.initials: B
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University Jinan, China.
authors.forename: Ruiqin
authors.initials: R
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University Jinan, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Feng
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University Jinan, China.
authors.forename: Fangzhong
authors.initials: F
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University Jinan, China.
authors.forename: Kuimei
authors.initials: K
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University Jinan, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University Jinan, China.
authors.forename: Kangle
authors.initials: K
authors.lastname: Niu
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University Jinan, China.
authors.forename: Quanquan
authors.initials: Q
authors.lastname: Yuan
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University Jinan, China.
authors.forename: Mingyu
authors.initials: M
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong UniversityJinan, China; State Key Laboratory of Microbial Technology, School of Life Science, Shandong University-Helmholtz Institute of Biotechnology, Shandong UniversityJinan, China.
authors.forename: Hailong
authors.initials: H
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong UniversityJinan, China; State Key Laboratory of Microbial Technology, School of Life Science, Shandong University-Helmholtz Institute of Biotechnology, Shandong UniversityJinan, China.
authors.forename: Youming
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University Jinan, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Fang
keywords: Cre-loxP system; Penicillium oxalicum; Tet-on system; doxycycline; selective markers