Nockemann S, Dlugonska H, Henrich B, Kitzerow A, Däubener W
Institute for Medical Microbiology and Virology, Düsseldorf, Germany.
Mol Biochem Parasitol. 1998 Nov 30;97(1-2):109-21. doi: 10.1016/s0166-6851(98)00138-8.
A Toxoplasma gondii tachyzoite expression library was screened with immune sera from T. gondii infected patients. Among others, one gene product reacted strongly with human sera and was further investigated. The gene called B10 was shown to encode a 41 kDa antigen. The complete genomic nucleotide sequence of the B10 protein has been analysed and was shown to contain one intron with conserved splice junctions. Southern blot analysis indicated that B10 is a single-copy gene. The corresponding 1.5 kb cDNA encodes a 318 amino acid sequence of mainly hydrophilic character with a putative signal sequence of 19 amino acids and no further trans-membrane domain. Immunofluorescence assays and immunoblots with a preparation of excreted-secreted antigens (ESA) suggested that the native protein is secreted into the parasitophorous vacuole and its delimiting membrane, indicating that B10 is a member of the ESA family of T. gondii. Recombinant B10 protein exhibited a strong reactivity with human serum samples both in ELISA and in immunoblots.
用来自弓形虫感染患者的免疫血清筛选了一个弓形虫速殖子表达文库。其中，一种基因产物与人类血清反应强烈，并对其进行了进一步研究。名为B10的基因被证明编码一种41 kDa的抗原。已对B10蛋白的完整基因组核苷酸序列进行了分析，结果显示其含有一个具有保守剪接位点的内含子。Southern印迹分析表明B10是一个单拷贝基因。相应的1.5 kb cDNA编码一个318个氨基酸的序列，该序列主要具有亲水性，有一个19个氨基酸的推定信号序列，且没有其他跨膜结构域。用排泄-分泌抗原（ESA）制剂进行的免疫荧光测定和免疫印迹表明，天然蛋白被分泌到寄生泡及其界定膜中，这表明B10是弓形虫ESA家族的一员。重组B10蛋白在ELISA和免疫印迹中均与人血清样本表现出强烈反应性。
