Takasuka T, Sayers N M, Anderson M J, Benbow E W, Denning D W
Department of Medicine, University of Manchester, UK.
FEMS Immunol Med Microbiol. 1999 Feb;23(2):125-33. doi: 10.1111/j.1574-695X.1999.tb01231.x.
The presence of catalases in the water soluble fractions of three Aspergillus fumigatus strains was investigated using non-denaturing and denaturing polyacrylamide gel electrophoresis and Western analysis. Using non-denaturing polyacrylamide gel electrophoresis and staining for catalase activity, three separate catalases were identified. An A. fumigatus catalase gene (catB) was cloned from genomic DNA using the Aspergillus niger catR gene as a probe. Polyclonal antibodies were raised to a glutathione S-transferase-CatB fusion product expressed in Escherichia coli. Western analysis indicated that, under denaturing conditions, the polyclonal antibody recognised a 90-kDa band and under non-denaturing conditions, two separate bands were identified. These results indicate that A. fumigatus in addition to CatB, produces at least two other catalases, one of which is similar in size to CatB. The polyclonal antibody was also used to observe catalase expression in mice, experimentally infected with A. fumigatus. Staining was observed heterogeneously throughout the fungal hyphae. This result indicates that catalase is produced by A. fumigatus during invasive aspergillosis.
运用非变性和变性聚丙烯酰胺凝胶电泳以及蛋白质免疫印迹分析，对三株烟曲霉菌株水溶性组分中过氧化氢酶的存在情况进行了研究。通过非变性聚丙烯酰胺凝胶电泳和过氧化氢酶活性染色，鉴定出三种不同的过氧化氢酶。以黑曲霉catR基因为探针，从基因组DNA中克隆出烟曲霉过氧化氢酶基因（catB）。针对在大肠杆菌中表达的谷胱甘肽S-转移酶-CatB融合产物制备了多克隆抗体。蛋白质免疫印迹分析表明，在变性条件下，多克隆抗体识别出一条90 kDa的条带，在非变性条件下，鉴定出两条不同的条带。这些结果表明，烟曲霉除了CatB外，还产生至少另外两种过氧化氢酶，其中一种的大小与CatB相似。该多克隆抗体还用于观察经烟曲霉实验性感染的小鼠体内过氧化氢酶的表达情况。在整个真菌菌丝中均观察到异质性染色。这一结果表明，在侵袭性曲霉病期间，烟曲霉会产生过氧化氢酶。
