AUTHOR=Jiang Baojie , Zhang Ruiqin , Feng Dan , Wang Fangzhong , Liu Kuimei , Jiang Yi , Niu Kangle , Yuan Quanquan , Wang Mingyu , Wang Hailong , Zhang Youming , Fang Xu 

TITLE=A Tet-on and Cre-loxP Based Genetic Engineering System for Convenient Recycling of Selection Markers in Penicillium oxalicum

JOURNAL=Frontiers in Microbiology

VOLUME=7

YEAR=2016

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2016.00485

DOI=10.3389/fmicb.2016.00485

ISSN=1664-302X

ABSTRACT=<p>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 <italic>Penicillium oxalicum</italic> JUA10-1(formerly named as <italic>Penicillium decumbens</italic>). To resolve this problem, we constructed a genetic manipulation system taking advantage of two established genetic systems: the Cre-<italic>lox</italic>P system and Tet-on system in <italic>P. oxalicum</italic> 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, <italic>ligD</italic> and <italic>bglI</italic>, were sequentially disrupted by <italic>lox</italic>P flanked <italic>ptrA</italic>. 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.</p>