Construction of the Aspergillus nidulans pmtC∆hogA∆yA∆ triple deletion strain by mating and benlate inducing

Tram Thi Huynh Le; Nhan Thy Le; Tien Thi Thuy Nguyen; Duong Thi Thuy Nguyen; Dong Xuan Nguyen

doi:10.46223/HCMCOUJS.tech.en.14.2.3208.2024

14(2)2024

Construction of the Aspergillus nidulans pmtC∆hogA∆yA∆ triple deletion strain by mating and benlate inducing

Author - Affiliation:

Tram Thi Huynh Le - Ho Chi Minh City University of Technology, Ho Chi Minh City Vietnam National University Ho Chi Minh City, Ho Chi Minh City , Vietnam

Nhan Thy Le - Nong Lam University of Hochiminh City, Ho Chi Minh City , Vietnam

Tien Thi Thuy Nguyen - Biotechnology Center of Hochiminh City, Ho Chi Minh City , Vietnam

Duong Thi Thuy Nguyen - Biotechnology Center of Hochiminh City, Ho Chi Minh City , Vietnam

Dong Xuan Nguyen - Biotechnology Center of Hochiminh City, Ho Chi Minh City , Vietnam

Corresponding author: Tram Thi Huynh Le - lthtram@hcmut.edu.vn

DOI:10.46223/HCMCOUJS.tech.en.14.2.3208.2024

Submitted: 24-01-2024
Accepted: 19-03-2024
Published: 04-09-2024

Abstract

In Aspergillus nidulans, construction of double or triple deletion strain is widely carried out to investigate genetic coordination, relation, or interaction. In this study, a fungal mutant, in which pmtC, hogA, and yA are deleted, is constructed for a recent experimental purpose. The mutation strain is screened and isolated by auxotrophic mutations from progenies of a sexual mating between the two A. nidulans fungi, pmtC∆yA∆ deletion strain and hogA∆ deletion strain. The genotype of the strain obtained from the matting is confirmed by PCR indicating that the obtained strain contains deleted hogA, both wild-type pmtC and deleted pmtC deletion. Moreover, the obtained strain forms morphology with irregular edges, abnormal growth, and derived colony, which is like general morphology of diploid strains, suggesting that the mutant is a diploid strain. As benlate is a haploidization-inducer for the diploid strain, the spores of the diploid strain are spread on media with 0.05ppm of benlate. Forming colonies performing pmtC-like characteristics, such as poor development, impaired vegetation and limited conidiation, are considered as target haploid strains and confirmed by PCR. The PCR results show that, among four isolated strains, there is only one strain showing PCR band which is approximate to pmtC knock-out gene, indicating the strain carry pmtC deletion gene.
As a result, the fungal strains, namely AnTLE36.HC, is a pmtC∆hogA∆yA∆ deletion strain.

Keywords

Aspergillus nidulans; benlate; mating; triple deletion

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Cite this paper as:

Le, T. T. H., Le, N. T., Nguyen, T. T. T., Nguyen, D. T. T., & Nguyen, D. X. (2024). Construction of the Aspergillus nidulans pmtC∆hogA∆yA∆ triple deletion strain by mating and benlate inducing. Ho Chi Minh City Open University Journal of Science – Engineering and Technology, 14(2), 28-38. doi:10.46223/HCMCOUJS.tech.en.14.2.3208.2024

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DOI: https://doi.org/10.46223/HCMCOUJS.tech.en.14.2.3208.2024