[Effect of sho1 gene of Aspergillus fumigatus on adaptation to osmotic pressure and on sensitivity of antifungal drugs].
OBJECTIVE
To construct the sho1 gene mutant of Aspergillus fumigatus, and to investigate the function of sho1 in adaptation to environmental changes.


METHODS
Saccharomyces cerivisiae sho1 gene homolog was identified by tblastn searches in A. fumigatus genome database. A PCR fragment, composed of the 5' flanking sequence (approximately 1 kb) of sho1, sho1 ORF, and its 3' flanking sequence (approximately 1 kb), was subcloned into a vector pDHt/SK2 with ApaI/XbaI enzyme to produce a recombinant plasmid PA. The selected marker pyrG cut from pLAX223 with NcoI/PstI was cloned into PA, and PB was produced. The plasmid PB was transformed into competent Agrobacterium tumefaciens EHA105 by using the freeze/thaw method. The strain of A. tumefaciens was designated as At sho1. Then triangle upsho1 was produced via ATMT. The radial growth was observed and compared between triangle up sho1 and wild type (wt) on MM, CM and BHI medium, and the sensitivity of triangle up sho1 and wt to amphotericin B, itraconazole, voriconazole, and caspofungin investigated.


RESULTS
Radial growth had visible difference between triangle up sho1 and wt on MM, CM and BHI medium. triangle up sho1 mutant was sensitive to high osmotic pressure but not to low osmotic pressure. The susceptibility to antifungal drugs was similar between the triangle up sho1 and wt.


CONCLUSIONS
Slow growth of the triangle up sho1 mutant has nothing to do with the different medium. The sho1 gene plays a role in response to osmotic pressure in A. fumigatus, but sho1 has a minor role in the susceptibility to the antifungal drugs despite the slow growth.
Department of Dermatology, Peking University First Hospital, Beijing 100034, China.
Yan
Y
Ma
Jian-jun
JJ
Qiao
Wei
W
Liu
Ruo-yu
RY
Li
D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000494:pharmacology*; D001232:Aspergillus fumigatus / Q000187:drug effects / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal; D008565:Membrane Proteins / Q000235:genetics*; D008826:Microbial Sensitivity Tests / Q000379:methods; D008969:Molecular Sequence Data; D009154:Mutation; D009997:Osmotic Pressure; D017385:Sequence Homology
D000935:Antifungal Agents; D005656:Fungal Proteins; D008565:Membrane Proteins