Puente Pilar, Leal Fernando
Departamento de Microbiología y Genética, Ed. Departamental Biología, Lab. 218, Paseo del Campo Charro s/n, Universidad de Salamanca, 37007 Salamanca, Spain.
Microbiology (Reading). 1998 Feb;144 ( Pt 2):561-567. doi: 10.1099/00221287-144-2-561.
A 996 bp Aspergillus nidulans cDNA encoding the ASPND1 immunodominant antigen was cloned and expressed in Escherichia coli as a fusion protein with the enzyme glutathione S-transferase (GST) from Schistosoma japonicum. The GST-ASPND1 fusion protein was purified from isolated bacterial inclusion bodies by preparative SDS-PAGE. After cleavage with thrombin, the ASPND1 recombinant antigen (ASPND1r) and the GST protein were separated by SDS-PAGE and immunoblotted with a number of different human sera. The sera from 22 (88%) of 25 patients with an aspergilloma recognized the ASPND1r recombinant antigen on immunoblots. Forty-nine normal human sera and 14 sera from patients with other infections were unreactive. The ASPND1r expressed in E. coli could therefore be used, in combination with previously reported recombinant antigens, as a standardized antigen for serological and clinical diagnosis of Aspergillus-associated diseases.
克隆了一段编码烟曲霉ASPND1免疫显性抗原的996 bp cDNA，并在大肠杆菌中表达为与日本血吸虫谷胱甘肽S-转移酶（GST）融合的蛋白。通过制备性SDS-PAGE从分离的细菌包涵体中纯化GST-ASPND1融合蛋白。用凝血酶切割后，通过SDS-PAGE分离ASPND1重组抗原（ASPND1r）和GST蛋白，并用多种不同的人血清进行免疫印迹。25例曲菌球患者中有22例（88%）的血清在免疫印迹中识别ASPND1r重组抗原。49份正常人血清和14份其他感染患者的血清无反应。因此，在大肠杆菌中表达的ASPND1r可与先前报道的重组抗原联合用作曲霉菌相关疾病血清学和临床诊断的标准化抗原。
