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Coccidioides posadasii Contains a Single 1,3-β-Glucan Synthase Gene That Appears To Be Essential for Growth

机译:球孢子虫含有一个单一的1,3-β-葡聚糖合酶基因,似乎对生长至关重要

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1,3-β-Glucan synthase is responsible for the synthesis of β-glucan, an essential cell wall structural component in most fungi. We sought to determine whether Coccidioides posadasii possesses genes homologous to known fungal FKS genes that encode the catalytic subunit of 1,3-β-glucan synthase. A single gene, designated FKS1, was identified, and examination of its predicted protein product showed a high degree of conservation with Fks proteins from other filamentous fungi. FKS1 is expressed at similar levels in mycelia and early spherulating cultures, and expression decreases as the spherules mature. We used Agrobacterium-mediated transformation to create strains that harbor ΔFKS1::hygB, a null allele of FKS1, and hypothesize that Fks1p function is essential, due to our inability to purify this allele away from a complementing wild-type FKS1 allele in a heterokaryotic strain. The heterokaryon appears normal with respect to growth rate and arthroconidium production; however, microscopic examination of strains with ΔFKS1::hygB alleles revealed abnormal swelling of hyphal elements.
机译:1,3-β-葡聚糖合酶负责β-葡聚糖的合成,β-葡聚糖是大多数真菌中必不可少的细胞壁结构成分。我们试图确定 Coccidioides posadasii 是否具有与已知真菌FKS基因同源的基因,这些基因编码1,3-β-葡聚糖合酶的催化亚基。鉴定出一个名为 FKS1 的单一基因,对其预测的蛋白质产物的检查显示,其与其他丝状真菌的Fks蛋白具有高度的保守性。 FKS1 在菌丝体和早期球状培养物中以相似的水平表达,并且随着球状体的成熟表达降低。我们用农杆菌介导的转化产生了带有ΔemsFKS1的无效等位基因Δ FKS1 :: hygB 的菌株。 ,并假设Fks1p功能是必不可少的,这是因为我们无法从异核生物株中的互补野生型 FKS1 等位基因中纯化出该等位基因。就生长速率和节肢动物的产量而言,异核体似乎是正常的。然而,对带有Δ FKS1 :: hygB 等位基因的菌株进行显微镜检查,发现其菌丝元素异常肿胀。

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