| Purification of the GfsA-3x FLAG Protein Expressed in Aspergillus nidulans
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Author:
Date:
2014-09-05
[Abstract] GfsA is a fungal β-galactofuranosyltransferase involved in the biosynthesis of O-glycan. To investigate the enzymatic functions of GfsA, we attempted to obtain a recombinant protein of this enzyme from two heterologous host organisms. However, GfsA could not be expressed as a recombinant protein in either Escherichia coli (E. coli) or Saccharomyces cerevisiae (S. cerevisiae). Therefore, we decided to employ Aspergillus nidulans (A. nidulans) as the host organism, and produced a strain that expressed 3x FLAG-tagged GfsA using ...
[摘要] GfsA是涉及O - 聚糖的生物合成的真菌β-半乳糖呋喃糖基转移酶。 为了研究GfsA的酶功能，我们尝试从两种异源宿主生物体获得该酶的重组蛋白。 然而，GfsA不能在大肠杆菌（大肠杆菌）或酿酒酵母（Saccharomyces cerevisiae）中表达为重组蛋白质（ cerevisiae ）。 因此，我们决定使用构巢曲霉（ A。nidulans ）作为宿主生物体，并产生使用染色体标签表达3×FLAG标记的GfsA的菌株。 为了证实其表达，从标记的菌株制备溶解的蛋白并用抗FLAG抗体分析。 表达3×FLAG标记的GfsA的菌株产生质量为约67kDa的功能性蛋白质。 该手稿中描述的方法允许纯化在A中表达的GfsA-3xFLAG蛋白。 构巢细胞。
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| Determination of D-galactofuranose Content of Galactomannoproteins in Aspergillus nidulans
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Author:
Date:
2014-09-05
[Abstract] Galactofuranose (Galf) is a component of several polysaccharides and glycoconjugates in certain species of filamentous fungi. Galf residues are frequently found in Aspergillus glycoproteins, including N-glycans and O-mannose glycans that modify many cell wall proteins and extracellular enzymes. It is known that furanoses, contained in oligosaccharides, are detected as pyranoses after hydrolysis, and that D-galactopyranose is not contained in the galactomannoproteins of Aspergillus spp. To determine the levels of D-galactofuranose in ...
[摘要] 半乳糖呋喃糖（Galactofuranose）（Gal ff）是某些种类的丝状真菌中几种多糖和糖缀合物的组分。 Gal< em>残基经常在曲霉属糖蛋白中发现，包括修饰许多蛋白质的聚糖和 细胞壁蛋白和细胞外酶。 已知包含在寡糖中的呋喃糖在水解后被检测为吡喃糖，并且D-吡喃半乳糖不包含在曲霉属的半乳糖蛋白中。 为了测定从构巢曲霉（构巢曲霉）提取的半乳糖蛋白中的D-半乳糖呋喃糖的水平，我们测量了半乳糖蛋白水解后D-吡喃半乳糖产生的量。 该手稿中描述的方法允许确定曲霉属中半乳糖甘露糖蛋白的D-半乳糖呋喃糖含量。
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