Ye X, Feng B, Szaniszlo P J
Division of Biological Sciences, The University of Texas at Austin, Austin, Texas 78712, USA.
Curr Genet. 1999 Oct;36(4):241-7. doi: 10.1007/s002940050496.
To explore potential virulence factors in the dematiaceous (melanized) fungus Wangiella dermatitidis, we established a gene expression system with properties of homologous transformation and color identification. Using a polyketide synthase gene (WdPKS1) fragment for targeting, we found that 52% of transformants became albinos easily distinguishable from nonspecific transformants. Southern analysis confirmed that the integrations were at the WdPKS1 locus, which however did not affect transformant growth. With a heterologous promoter, P-glaA, enhanced expression of lacZ was found at 37 degrees C. Our results indicated that this system allows the efficient production of isogenic strains for gene function analysis in W. dermatitidis.
为了探索暗色(黑色素化)真菌皮炎万吉拉霉(Wangiella dermatitidis)中的潜在毒力因子,我们建立了一个具有同源转化和颜色鉴定特性的基因表达系统。使用聚酮合酶基因(WdPKS1)片段作为靶向,我们发现52%的转化体变成白化体,很容易与非特异性转化体区分开来。Southern分析证实整合发生在WdPKS1基因座,然而这并不影响转化体的生长。使用异源启动子P-glaA,在37℃时发现lacZ的表达增强。我们的结果表明,该系统能够有效地产生同基因菌株,用于皮炎万吉拉霉的基因功能分析。