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Volume 8
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Article

Establishment of High-Efficiency Screening System for Gene Deletion in Fusarium venenatum TB01

by
Sheng Tong
1,2,
Kexin An
1,2,
Wenyuan Zhou
1,2,
Wuxi Chen
1,2,
Yuanxia Sun
1,2,
Qinhong Wang
1,2 and
Demao Li
1,2,*
1
Tianjin Key Laboratory for Industrial Biological Systems and Bioprocessing Engineering, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China
2
National Center of Technology Innovation for Synthetic Biology, Tianjin 300308, China
*
Author to whom correspondence should be addressed.
J. Fungi 2022, 8(2), 169; https://doi.org/10.3390/jof8020169
Submission received: 14 January 2022 / Revised: 4 February 2022 / Accepted: 5 February 2022 / Published: 10 February 2022
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Abstract

Genetic engineering is one of the most effective methods to obtain fungus strains with desirable traits. However, in some filamentous fungi, targeted gene deletion transformant screening on primary transformation plates is time-consuming and laborious due to a relatively low rate of homologous recombination. A strategy that compensates for the low recombination rate by improving screening efficiency was performed in F. venenatum TB01. In this study, the visualized gene deletion system that could easily distinguish the fluorescent randomly inserted and nonfluorescent putative deletion transformants using green fluorescence protein (GFP) as the marker and a hand-held lamp as the tool was developed. Compared to direct polymerase chain reaction (PCR) screening, the screening efficiency of gene deletion transformants in this system was increased approximately fourfold. The visualized gene deletion system developed here provides a viable method with convenience, high efficiency, and low cost for reaping gene deletion transformants from species with low recombination rates.
Keywords: Fusarium venenatum; homologous recombination rate; screening efficiency; visualized gene deletion system; fluorescence observation; ku70 Fusarium venenatum; homologous recombination rate; screening efficiency; visualized gene deletion system; fluorescence observation; ku70

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MDPI and ACS Style

Tong, S.; An, K.; Zhou, W.; Chen, W.; Sun, Y.; Wang, Q.; Li, D. Establishment of High-Efficiency Screening System for Gene Deletion in Fusarium venenatum TB01. J. Fungi 2022, 8, 169. https://doi.org/10.3390/jof8020169

AMA Style

Tong S, An K, Zhou W, Chen W, Sun Y, Wang Q, Li D. Establishment of High-Efficiency Screening System for Gene Deletion in Fusarium venenatum TB01. Journal of Fungi. 2022; 8(2):169. https://doi.org/10.3390/jof8020169

Chicago/Turabian Style

Tong, Sheng, Kexin An, Wenyuan Zhou, Wuxi Chen, Yuanxia Sun, Qinhong Wang, and Demao Li. 2022. "Establishment of High-Efficiency Screening System for Gene Deletion in Fusarium venenatum TB01" Journal of Fungi 8, no. 2: 169. https://doi.org/10.3390/jof8020169

APA Style

Tong, S., An, K., Zhou, W., Chen, W., Sun, Y., Wang, Q., & Li, D. (2022). Establishment of High-Efficiency Screening System for Gene Deletion in Fusarium venenatum TB01. Journal of Fungi, 8(2), 169. https://doi.org/10.3390/jof8020169

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