Chow C, Gauci C G, Cowman A F, Lightowlers M W
Veterinary Clinical Centre, University of Melbourne, Princes Highway, Werribee, Victoria 3030, Australia.
Mol Biochem Parasitol. 2001 Nov;118(1):83-8. doi: 10.1016/s0166-6851(01)00373-5.
Echinococcus granulosus causes cystic hydatidosis in humans. A recombinant antigen vaccine has been developed, for use in the parasite's natural animal intermediate hosts, that may provide a new tool for control of hydatid disease transmission. The antigen, designated EG95, is encoded by a cDNA the features of which indicate it to be an incomplete copy of the associated mRNA. Characterisation of the gene(s) encoding the antigen was undertaken in order to enable subsequent study of genetic variability in the gene and associated protein in different parasite isolates. Southern hybridisation studies of E. granulosus genomic DNA probed with the eg95 cDNA revealed that the gene belonged to a gene family. DNA sequence analysis of cloned genomic fragments indicated that the gene family consists of at least seven members, one of which is a pseudogene. The gene having identity with the eg95 cDNA was cloned and sequenced, and the full length mRNA characterised. Genomic sequence and structure of the eg95 gene family members are highly conserved with respect to the gene encoding EG95. Four eg95-related genes are predicted to express an identical EG95 protein and all four were shown to be expressed in the oncosphere life-cycle stage. The full length EG95 protein has a predicted molecular mass of 16.9 kDa, secretory signal sequence, carboxy-terminal glycosylphosphatidylinositol hydrophobic anchor motif and a fibronectin type III domain. PCR amplification conditions were established which allow gene-specific characterisation of the eg95 gene in E. granulosus isolates from different host species and geographical locations.
细粒棘球绦虫可导致人类患囊型包虫病。已开发出一种重组抗原疫苗,用于该寄生虫的天然动物中间宿主,这可能为控制包虫病传播提供一种新工具。该抗原被命名为EG95,由一个cDNA编码,其特征表明它是相关mRNA的不完全拷贝。对编码该抗原的基因进行了表征,以便随后研究不同寄生虫分离株中该基因及相关蛋白质的遗传变异性。用eg95 cDNA探测细粒棘球绦虫基因组DNA的Southern杂交研究表明该基因属于一个基因家族。对克隆的基因组片段进行DNA序列分析表明该基因家族至少由七个成员组成,其中一个是假基因。克隆并测序了与eg95 cDNA具有同一性的基因,并对全长mRNA进行了表征。eg95基因家族成员的基因组序列和结构在编码EG95的基因方面高度保守。预测有四个与eg95相关的基因表达相同的EG95蛋白,并且已证明所有四个基因在六钩蚴生命周期阶段均有表达。全长EG95蛋白的预测分子量为16.9 kDa,具有分泌信号序列、羧基末端糖基磷脂酰肌醇疏水锚定基序和一个纤连蛋白III型结构域。建立了PCR扩增条件,可对来自不同宿主物种和地理位置的细粒棘球绦虫分离株中的eg95基因进行基因特异性表征。