A Tet-on and Cre-loxP Based Genetic Engineering System for Convenient Recycling of Selection Markersin Penicillium oxalicum

作者：Jiang, Baojie; Zhang, Ruiqin; Feng, Dan; Wang, Fangzhong; Liu, Kuimei; Jiang, Yi; Niu, Kangle; Yuan, Quanquan; Wang, Mingyu; Wang, Hailong; Zhang, Youming; Fang, Xu^*

来源：Frontiers in Microbiology, 2016, 7: 485.

DOI：10.3389/fmicb.2016.00485

• selective markers
• Penicillium oxalicum
• Cre-loxP system
• Tet-on system
• doxycycline

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. Introduction

• 出版日期2016-4-12
• 单位微生物技术国家重点实验室; 山东大学